GB2128986A

GB2128986A - Derivatives of 6 b-hydroxalkylpenicillanic acids as b-lactamase inhibitors

Info

Publication number: GB2128986A
Application number: GB08316829A
Authority: GB
Inventors: Michael Stephen Kellogg
Original assignee: Pfizer Inc
Current assignee: Pfizer Inc
Priority date: 1979-10-22
Filing date: 1983-06-21
Publication date: 1984-05-10
Also published as: DD154542A5; RO86176A; NL8005810A; NO159019B; NL183358B; PL235907A1; IL61307A0; AR227164A1; GB8316830D0; ES496166A0; PT71946B; IE850293L; RO81225B; NZ195307A; FI71156C; DE3039504C2; RO81225A; BE885812A; JPS5665892A; MX6094E

Description

1

SPECIFICATION

Derivatives of 60-hydroxyalkylpenicillanic acids as P-lactamase inhibitors GB 2 128 986 A 1 Background of the invention

One of the most well-known and widely used class of antibacterial agents are the so-called P-lactam antibiotics. These compounds are characterized in that they have a nucleus consisting of a 2-azeticlinone (p-lactam) ring fused to either a thiazolicline or a dihydro-1,3-thiazine ring. When the nucleus contains a thiazolidine ring, the compounds are usually referred to generically as penicillins, whereas when the nucleus contains a dihydrothiazine ring, the compounds are referred to as cephalosporins. Typical examples of penicillins which are commonly used in clinical practice are benzy1penicillin (penicillin G), phenoxymethyl penicillin (penicillin V), ampicillin and carbenicillin; typical examples of common cephalosporins are cephalothin, cephalexin and cefazolin.

However, despite the wide use and wide acceptance of the p-lactam antibiotics as valuable chemother apeutic agents, they suffer from the major drawback that certain members are not active against certain microorganisms. It is thought that in many instances this resistance of a particular microorganism to a given p-lactam antibiotic results because the microorganism produces a p- lactamase. The latter substances are enzymes which cleave the p-lactam ring of penicillins and cephalosporins to give products which are devoid of antibacterial activity. However, certain substances have the ability to inhibit P-lactamases, and when a P-lactamase inhibitor is used in combination with a pencillin or cephaslosporin it can increase or enhance 20 the antibacterial effectiveness of the penicillin or cephalosporin against certain P-lactamase producing microorganisms. It is considered that there is an enhancement of antibacterial effectiveness when the antibacterial activity of a combination of P-lactamase inhibiting substance and a P-lactam antibiotic is significantly greaterthan the sum of the antibacterial activities of the individual components against P-lactamase producing microorganisms.

The present invention relates to a series of 6p-hydroxyalkylpenicillanic acids and esters thereof readily hydrolyzable in vivo which are potent inhibitors of microbial P- lactamases and which enhance the effectiveness of p-lactam antibiotics. The invention further relates to benzyl 6p-hydroxyalkyl pen icillanates, said esters being useful chemical intermediates to the corresponding acids.

Pharmaceutical compositions comprising the above-mentioned 6p-substituted penicillanic acids and readily hydrolyzable esters thereof with certain plactam antibiotics as well as a method for increasing the effectiveness of certain P-lactam antibiotics in combination with the above-mentioned 6p-substituted penicillanic acids and readily hydrolyzable esters thereof are also part of the present invention.

Di Ninno, et. al., [J. Org. Chem., 42, 2960 (1977)] have reported the synthesis of 6p-hydroxyalkylpenicillanic 0.5 acids and the corresponding benzyl esters as potential antibacterial agents and useful intermediates, respectively.

6-Ethylpenicillanic acid and its sulfoxide derivative are claimed as antibiotics in U.S. Patent 4,123,539.

6ot-Hydroxypenicillanic acid and esters thereof have been prepared from 6diazopenicillanic acid and the corresponding esters [J. Org. Chem., 39 1444 (1974)].

U.S. Patent 4,143,046 discloses 6p-substituted sulfonyloxypenicillanic acids as antibacterial agents. 40 Summary of the invention

The compounds of this invention are of the formulae H H H H 02 45 CH CH S 3 3 R CH 3 CH 3 0 C0 2 R 1 0 C0 2 R 1 1 and 11 50 or a pharmaceutically acceptable base salt thereof, wherein R is a I kylsu Ifo nyloxym ethyl having one to four carbon atoms in the alkyl group, phenyl su Ifo nyl oxym ethyl, substituted phenylsulfonyloxymethyl (wherein said substituentis methyl, methoxy,fluoro,chloro, bromo ortrifluoromethyl) or 1-hydroxy-3-phenylpropyl; 56 R, is benzyl, hydrogen or ester-forming residues readily hydrolyzable in vivo; and R2 is O-R3 I R4-L;11- wherein R3 is sulfo, hydrogen, alkoxycarbonyl of two to four carbon atoms, alkanoyl of two to eighteen carbon atoms, benzoyi, substituted benzoyl (wherein said substituent is amino, methyl, methoxy, fluoro, chloro, bromo or trifluoromethyl), alkylsulfonyl of one to four carbon atoms, phenyisuifonyl or substituted phenyIsulfonyl (wherein said substituent is methyl, methoxy, fluoro, chloro, bromo or trifluoromethyl); and R4 is hydrogen, alkyl of one to four carbon atoms, phenyl, benzyi, pyridyl or p-phenethyl.

2 GB 2 128 986 A A preferred group of P-lactamase inhibitors are those of formula 11 wherein R, and R4 are each hydrogeh. Especially preferred within this group are those compounds wherein R3 is hydrogen, acetyl, stearoyl or benzoyl.

A second group of preferred compounds related to 11 are those wherein R, and R3 are each hydrogen and R4 is alkyl of one to four carbon atoms. Especially preferred within this group are the compounds wherein R4 5 is methyl.

Athird group of preferred compounds of formula I are those wherein R, is hydrogen. Especially preferred within this group are those compounds wherein R is m ethylsu Ifo nyl oxym ethyl, p-toluenesulfonyloxymethyl and 1-hydroxy-3-phenylpropyl.

The present invention also relates to a pharmaceutical composition useful for treating bacterial infections 10 in mammals, which comprises a pharmaceutically acceptable carrier, a P-lactam antibiotic and a compound selected from the formulae R2 is 2 H H H H 0 2 CH CH S 3 S 3 15 R lh CH 3 R 2 CH3 Co 2RS 0 CO 2 R 5 and IV 20 or a pharmaceutically acceptable base salt thereof, wherein R is alkylsulfonyloxymethyl having one to four carbon atoms in the alkyl group, ph enylsu Ifo nyl oxym ethyl, substituted ph enylsu Ifo nyl oxym ethyl wherein said substituent is methyl, methoxy, fluoro, chloro, bromo ortrifluoromethyl or 1-hydroxy-3-phenylpropyl; O-R3 1 R4-(ll- wherein R3 is sulfo, hydrogen, alkoxycarbonyl of two to four carbon atoms, alkanoyl of two to eighteen carbon atoms, benzoyl, substituted benzoyl wherein said substituent is amino, methyl, methoxy, fluoro, chloro, bromo or trifluoromethyl, alkylsulfonyl of one to four carbon atoms, phenylsulfonyl or substituted phenylsulfonyl wherein said substitutent is methyl, methoxy, fluoro, chloro, bromo or trifluoromethyl; R4 is hydrogen, alkyl of one to four carbon atoms, phenyl, pyridyl, benzyl or p- phenethyl; and R5 is hydrogen or 35 ester-forming residue readily hydrolyzable in vivo.

Preferred compounds are those of formulae III and IV wherein R5 is hydrogen or ester-forming residues readily hydrolyzable in vivo selected from alkanoyloxymethyl of three to six carbon atoms, 1 (alkanoyloxy)ethyl of four to seven carbon atoms, 1 -methyl-1 -(alkanoyloxy)ethyl of five to eight carbon atoms, alkoxycarbonyloxymethyl of three to six carbon atoms, 1-(alkoxycarbonyloxy)ethyl of fourto seven 40 carbon atoms, 1 -methyl-1 (alkoxycarbonyloxy)ethyl of five to eight carbon atoms, 3-phthalidyl, 4crotonolactonyl, and --butyrolacton-4-yl, and said P-lactam antibiotics are selected from penicillins and cephalosporins. Especially preferred are compounds of formula IV wherein R4 and REj are each hydrogen and R3 is hydrogen, acetyl, stearoyl or benzoyl. Also especially preferred is the component wherein R3 and R5 are each hydrogen and R4 is methyl. Especially preferred are compounds of formula III wherein R is 45 methylsu Ifo nyl oxym ethyl p-to I uen esu Ifo nyloxym ethyl and 1 -hyd roxy-3-p henyl propyl.

The invention also consists of a method for increasing the effectiveness of a P-lactam antibiotic in a mammalian subject which comprises coadministration to said subject a p-lactam antibiotic effectiveness increasing amount of a compound of the formulae H H H H 0 2 CH 3 50 CH S..

R k CH 3 R 2 On CH 3 3 CO R 0 CO R 2 5 2 5 and IV 55 or a pharmaceutically acceptable base salt thereof wherein R is alkylsulfonyloxymethyl having one to four carbon atoms in the alkyl group, ph enylsu Ifo nyl oxym ethyl, substituted phenylsulfonyloxymethyl wherein said substituent is methyl, methoxy, fluoro, chloro, bromo ortrifluoromethyl or 1-hydroxy-3-phenylpropyl; R2 is O-R3 1 R4-L;M_ wherein R3 is sulfo, hydrogen, alkoxycarbonyl of two to four carbon atoms, alkanoyl of two to eighteen 65 3 GB 2 128 986 A 3 carbon atoms, benzoyl, substituted benzoyl wherein said substituent is amino, methyl, methoxy, fluoro, chloro, bromo ortrifluoromethyl, alkylsulfonyl of one to four carbon atoms, phenylsulfonyl or substituted phenylsulfonyl wherein said substituent is methyl, methoxy, fluoro, chloro, bromo ortrifluoromethyl and R4 is hydrogen, alkyl of one to four carbon atoms, phenyl, benzyl, pyridyl or P-phenethyl; and R5 is hydrogen or 5 ester-forming residue readily hydrolyzable in vivo. Preferred compounds are those of formula III and IV wherein R5 is hydrogen or ester- forming residues readily hydrolyzable in vivo selected from alkanoyloxymethyl having from 3 to 6 carbon atoms, 1-(alkanoyloxy)-ethyl having from 4 to 7 carbon atoms, 1-methyl-l-(alkanoyloxy)ethyl having from 5 to 8 carbon atoms, alkoxycarbonyloxymethyl having from 3 to 6 carbon atoms, 1-(alkoxycarbonyloxy)ethyl having from 4 to 7 carbon atoms, 1-methyl-l-(alkoxycarbonyloxy)ethyI having from 5 to 8 carbon atoms, 10 3-phthalidyl, 4-crotonolactonyl andy-butyrolacton-4-yl, and said p-lactam antibiotics are selected from penicillins and cephalosphorins. Especially preferred are compounds related to IV wherein R4 and R5 are each hydrogen and R3 is hydrogen, acetyl, stearoyl or benzoyl. Also especially preferred of this class is the compound wherein R3 and RF, are each hydrogen and R4 is methyl.

Especially preferred are compounds of formula I H wherein R ism ethylsu Ifo nyl oxym ethyl, ptoluenesulfonyloxymethyl and 1-hydroxy-3phenylpropyl. The definition of the aforementioned ester-forming residues is meant to embrace any ester which is readily hydrolyzed in a mammal. Especially preferred are esterforming residues readily hydrolyzable in vivo selected from alkanoyloxymethyl of three to six carbon atoms, 1(alkanoyloxy)ethyl of four to seven carbon atoms, 1-methyl-l-(alkanoyloxy)ethyl of five to eight carbon atoms, a I koxyca rbo nyloxym ethyl of three to six 20 carbon atoms, 1(alkoxycarbonyloxy)ethyl of fourto seven carbon atoms, 1-methyl1(alkoxycarbonyloxy)ethyl of five to eight carbon atoms, 3-phthalidyl, 4crotonolactonyl, and -y-butyrolacton4-yl. The preferred P-lactams whose antibiotic activity is enhanced by the 6p-hydroxyalkylpenicillanic acid sulfones of the present invention are selected from:

6-(2-phenylacetamido)penicillanic acid, 6-(2-phenoxyacetamido)penicillanic acid, 6-(2-phenyl propionam ido)penicil Ian ic acid, 6-(D-2-a mi no-2-phenylaceta m ido) penici Ilan ic acid, 6-(D-2-amino-2-[4-hydroxyphenyl]acetamido)penicillanic acid, 6-(D-2-amino-2-[1,4-cyclohexadienyl]acetamido)penicillanic acid 6-(l-aminocyclohexanecarboxamido)penicillanic acid, 6-(2-carboxy-2-phenylacetamido)penicillanic acid, 6-(2-carboxy-2-[3-thienyllacetamido)penicillanic acid, 6-(D-2-[4-ethylpiperazin-2,3-dione-l-carboxamido]-2phenylacetamido)penicill anic acid, 6-(D-2-[4-hydroxy-1,5-na phthyridi ne-3-carboxa m idol-2- phenylacetamido)pen icil Ian ic acid, 6-(D-2-sulfo-2-phenylacetamido)penicillanic acid, 6-(D-su Ifoa m ino-2-phenylacetam ido) pen ici Ilan ic acid, 6-(D-2-[imidazolidin-2-one-l-carboxamidol-2-phenylacetamido)penicillanic acid 6-(D-2-[3-methylsu Ifonyl i midazol idi n-2-one-1 -carboxamidol-2-phenyl aceta mido) pen icil Ian ic acid, 40 6-([hexahydro-1 H-azepin-1-yllmethyleneamino)penicillanic acid, acetoxymethyl 6-(2-phenylacetamido)penicillanate, acetoxymethyl 6-(D-2-amino-2-phenylacetamido)penicillanate, acetoxymethyl 6-(D-2-amino-2-[4-hydroxyphenyllacetamido)penicillanate, pivaloyloxymethyl 6-(2-phenylacetamido)penicillanate, piva loyl oxym ethyl 6-(D-2-amino-2-phenylacetamido)penicillanate, pivaloyloxymethyl 6-(D-2-am ino-2-[4-hyd roxyphenyl Jacetarn ido) penici Ila nate, 1 -(ethoxyca rbo nyl oxy) ethyl 6-(2-phenylacetamido)penicillanate,l(ethoxycarbonyloxy)ethyI 6-(D-2 amino-2-phenylacetamido)pencillanate, 1 -(eth oxyca rbo nyl oxy) ethyl 6-(D-2-amino-2-[4hydroxyphenyllacetamido)penicillanate, 3-phthalidyl 6-(2-phenylaceta mido) pen icil lanate, 3-phthalidyl 6-(D-2-amino-2phenylacetamido)penicillanate, 3-phthalidyl 6-(D-2-amino-2-[4hydroxyphenyllacetamido)penicillanate, 6-(2-phenoxycarbonyl-2phenylacetamido)penicillanic acid, 55 6-(2-tolyloxycarbonyl-2phenylacetamido)penicillanic acid, 6-(2-[5-indanyloxycarbonyll-2phenylacetamido)penicillanic acid 6-(2-phenoxycarbonyl-2-[3thienyl]acetamido)penicillanic acid, 6-(2-tolyloxycarbonyl-2-[3thienyllacetamido)penicillanic acid, 6-(2-[5-indanyloxycarbonyl]-2-[3thienyllacetamido)penicillanic acid, 60 6-(2,2-dimethyl-5-oxo-4-phenyl-1 imidazolidinyl)penicillanic acid, 7-(2-[2thienyllacetamido)cephalosporanic acid,, 7-(2-[l-tetrazolyl]acetamido-3(2-[5-methyl-1,3,4-thiadiozolyllthiomethyl)-3 desacetoxymethy1cephalosporanic acid, 7-(D-2-formyloxy-2-phenylacetamido)3-(5-[l-methyltetrazolyf]thiomethyl)-365 desacetoxymethy1cephalosporanic acid, 4 GB 2 128 986 A 7-(D-2-amino-2-phenylacetamido)desacetoxycephalosporanic acid, 7-alphamethoxy-7-(2-[2-thienyllacetamido)-3-carbamoyloxymethyl-3-desacetoxy methylcephalosporanic acid, 7-(2-cyanoacetamido)cephalosporanic acid, 7(D-2-hydroxy-2-phenylacetamido)-3-(5-[l-methyltetrazolyllthiomethyl)3desacetoxymethy1cephalosporanic acid, 7-(D-2-amino-2-phydroxypehnylacetamido)desacetoxycephalosporanic acid, 7-(2-[4pyridylthiolactamido)cephalosporanic acid,7-(D-2-amino-2[1, 4cyclohexadienyllactamido)cephalosporanic acid, 7-(D-2-amino-2-phenylacetamido)cephalosporanic acid, 7-[D-(-)-alpha-(4ethyl-2,3-dioxo-1 -piperazinecarboxamido)-alpha-(4hydroxyphenyl)acetamido]-3-[(lmethyl-1,2-3,4-tetrazol-5-yl)thiomethyll-3cephem-4-carboxylic acid, 7-(D-2-amino-2-phenylacetamido)-3-chloro-3cephem-4-carboxylic acid, [2-(2-amino-4-thiazolyl)-2(methoximino)acetamidolcephalosporanic acid, [6R,7R-3-carbamoyloxymethyl-7(2Z)-2-methoxyimino(fur-2-yl)acetamido-ceph3-e m-4-carboxylate] 15 7-(2-(2-aminothiazol-4-yl)actamido]-3-[([1-2-dimethylaminoethyl)-I Htetrazo 1-5-yl Ith io) m ethyl Iceph-3-em4-carboxylic acid, and a pharmaceutically acceptable salts thereof.

4 Detailed description of the invention

The p-lactamase inhibitors of the present invention are conveniently prepared starting with benzyl 20 6,6-dibromopenicillanate. The condensation of an appropriate aldehyde with the enolate formed through the reaction of benzyl 6,6-dibromopenicillanate with an organometallic reagent, such as the process taught by Di Ninno, et. aL, J. Org. Chem., 42,2960 (1977), results in the formation of a benzyl 6-bromo-6 hydroxyalkylpenicillanate, the initial intermediate leading to the products of the present invention.

The product of this initial condensation is comprised of diastereomeric mixtures due to two asymmetric 25 centers, one at the 6-position of the penam nucleus and the second at the carbon atom in the chain at the 6-position, shown as follows:

H Ho Br S CH 3 CH 3 R4 3...C0 2 CH 2 c 6 H 5 As one skilled in the art can readily determine, of these only one is an asymmetric center in this intermediate product when R4 is hydrogen.

The substituents at the 6-position are designated as at or 0 and are so indicated in the structural formula by a broken or solid bond, respectively. The stereochemical configuration of the substituent in the side chain is designated as (R) or (S) (Cahn, etal., Experientia, 12, 81 (1956). The assignment of configuration is based on 40 nuclear-magnetic-resonance spectroscopy.

Experimentally, benzyi 6,6-dibromopenicillanate in a reaction-inert solvent at -20 to -78'C. is treated with about one equivalent of t-butyl lithium or t-butyl magnesium chloride. The resulting enolate is then treated with the appropriate aldehyde and, after a short reaction period, the reaction is quenched and the product isolated by conventional means.

The addition of zinc chloride to a solution of the enolate prior to the addition of aldehyde appears to exert control over the stereochemistry of the condensation product. Accordingly, a high preponderance of (S) configuration in the side chain is obtained under these conditions.

When diethyl zinc is employed as the initial organometallic reagent a preponderance of (R) configuration in the side chain of the product is obtained.

The initial reaction is conducted in an anhydrous reaction-inert solvent, which appreciably solubilizes the reactants without reacting to any great extent with the reactants or products under reaction conditions. It is preferred that said solvents have boiling points and freezing points compatible with reaction temperatures. Such solvents or mixtures thereof include aromatic solvents such as toluene and ethereal solvents such as tetrahydrofuran and diethyl ether.

The molar ratio of the starting penicillanate derivative and the organometallic reagent is not critical to the process. The use of a slight excess of organometallic, up to as much as a ten percent above an equimolar quantity, aids in the completion of the reaction and offers no serious problems in isolating the desired product in purified form.

Moisture can effectively be kept out of the reaction by employing a nitrogen or argon atmosphere. 60 Reaction time is inherently dependent on concentration, reaction temperature and reactivity of the starting reagents. When the reaction is conducted at the preferred reaction temperatue of -60 to -78'C. the reaction time for the formation of the enolate is about 30-45 minutes. The reaction time forthe formation of the intermediate product from the aforementioned enolate and aldehyde is about 30-60 minutes.

On completion of the reaction, the product is isolated by conventional means and the diastereomeric GB 2 128 986 A 5 mixture can be separated by column chromatography. However, the nature of the next reaction, which is the removal of the 6-bromo substituent, precludes the necessity for said separation of ot and p epimers at C-6.

Treatment of the benzyl 6-bromo-6-hydroxyalkylpenicillanate, resulting from the first reaction, with tri-n-butyltin hydride leads to the formation of a benzyi 6-hydroxyalkylpenicillanate in which the 6-hydroxyalkyl moiety is in the P-conf igu ration. This result is independent of the conformation of the substituents at the 6-position of the starting regions. Thus 6a-bromo-6p- hydroxyalkyl esters and 6p-bromo-6a.-hydroxyalkyl esters both give, on treatment with tri-nbutyltin hydride, the same 6p hydroxyalkyl ester as the main product, assuming all other structural parameters in the compounds are the same.

The reaction is carried out in a reaction-inert solvent which appreciably solubilizes the reactants without 10 reacting to any great extent with the reactants or the product under reaction conditions. It is further preferred that said solvent be an aprotic solvent, immiscible with water and have a boiling and freezing point compatible with reaction temperatures. Such solvents or mixtures thereof include the preferred solvents benzene and toluene.

Reaction time is dependent on concentration, reaction temperature and reactivity of the reagents. When 15 the reaction is carried out at the preferred temperature, the reflux temperature of the solvent, the reaction is usually complete in about 4-5 hours.

The molar ratio of the reagents is not critical to the process. Usually an excess of the tin hydride is employed and as much as a 100% excess over an equimolar amount can be employed.

When the reaction is complete the solvent is removed and the residue triturated with hexane to remove 20 the organotin by-product. The intermediate product can be purified and the isomers separated by column chromatography.

The oxidation of the resulting benzyl 6p-(S) and (R)hydroxyal kyl pen icil lanate to the corresponding sulfones of formula 11 wherein R, is benzyi is conveniently carried out using an organic peroxy acid, e.g., a peroxycarboxylic acid such as m-chloroperbenzoic acid. The reaction is carried out by treating the 25 appropriate benzy] 6P-(R) or (S)hydoxyikylpenicillanate with about 2 to about 4 equivalents and preferably about 2.2 equivalents of the oxidant in a reaction-inert solvent. Typical solvents are chlorinated hydrocarbons, such as methylene chloride, chloroform and 1,2- dichloroethane, The oxidant and substrate are initially combined in a solvent at O-YC. The temperature is allowed to rise to room temperature. Reaction time is about 3-6 hours.

During isolation of the sulfones, which are useful intermediates, the solvent is removed and the residue partitioned between water and a water immiscible solvent such as ethyl acetate. The pH of the water-organic solvent mixture is adjusted to 7.0 and any excess peroxide is decomposed with sodium bisulfite. The intermediate product, which is contained in the organic phase, is isolated and purified by conventional means.

The biol ogically- active products of the present invention of formula 1 and 11 wherein R, is hydrogen are prepared by debenzy[ation of the corresponding benzyl esters. Accordingly, the appropriate benzyl ester is added to a suspension of prehydrogenated 5% palladium-on-calcium carbonate catalyst in a 50% methanol-water solution. The hydrogenolysis is conducted at room temperature and is usually conducted at 45-50 psi pressure. Under these conditions the reaction is usually complete in 30-60 minutes. Filtration of the 40 spent catalyst followed by removal of the solvent by freeze drying results in the isolation of the calcium salt.

Acidification of the the filtrate, after removal of the catalyst, followed by extraction with a water immiscible solvent such as ethyl acetate, results in isolation of the free acid wherein R, is hydrogen.

Alternatively, the compounds of formula 11 wherein R, is hydrogen can also be prepared by the same series of reactions previously described, but in a different sequential order. For example, the initially formed 45 benzyi 6-bromo-6-hyd roxya lkyl pen icil lanates can be oxidized as previously described followed by removal of the 6-bromo substituents with tri-n-butyltin hydride and debenzylation.

Compounds of the present invention of formula 11 wherein R3 is alkanoyl are prepared by acylation of the requisite benzyl 6phydroxyalkylpenicillanate sulfone followed by hydrogenoiysis of the benzyi ester to give those compounds of formula 11 wherein R3 is as indicated and R, is hydrogen.

The acylation is carried out by contacting the appropriate pencillanate sulfone with an equimolar amount of the requisite acid halide, plus as much as a 10-20% excess, in a reaction-inert solvent such as methylene chloride. A tertiary amine, added in molar amounts corresponding to the acid halide, acts as a scavenger for the hydrogen halide formed.

The acylation is conducted at a reaction temperature of about 0-5'C. and requires a reaction time of about 55 20-30 minutes. The intermediate product is obtained by removing the solvent and treating the resulting residue with water-ethyl acetate and the organic phase is evaporated to give the desired materials.

The formation of the final products, compounds of formula 11 wherein R3 is alkanoyl and R, is hydrogen, is carried out by debenzylation under conditions hereinbefore described.

The compounds of the present invention of formula 11 wherein R3 is benzoyf, said substituted benzoyi, or 60 said alkoxycarbonyl are all prepared in a manner similar to the preparation of those compounds wherein R3 is alkanoyl, and comprises initial acylation of the appropriate benzyl 6p- hydroxyalkylpenicillanate sulfone followed by removal of the benzyl moiety from ester.

The compounds of the present invention of formula [wherein R is said alky[suifonyloxymethyi, phenyisufonlyoxymethyl or said substituted phenyisuifonyloxymethyl and of formula 11 wherein R3 is said 65 6 GB 2 128 986 A 6 alkylsulfonyl, phenylsulfonyl or said substituted phenylsulfonyl are most conveniently prepared by the initial acylation of the requisite benzyl 6p- hydroxylalkylpenicillanate with about an equi-molar amount of the appropriate sulfonyl chloride, employing pyridine as the solvent and at a reaction temperature of O'C. and reaction time of about 2-3 hours. The product is isolated by quenching of the reaction mixture with water followed by extraction and purification.

The second reaction in the series comprises debenzylation of the intermediate ester using hydrogen and 5% palladium-on-calcium carbonate, a procedure previously described.

The final step to compounds of formula 11 comprises oxidation of the 6psulfonyloxyalkylpenicillanic acids with potassium permanganate in a mixture of water-methylene chloride at ambient temperatures at a pH of 6-6.4. Following the reaction, which requires about 45-60 minutes, the pH is adjusted to 1.5 and the product 10 isolated from the organic phase.

The synthesis of those compounds of formula 11 wherein R3 is SUIfO is effected by the reaction of the appropriate benzyl 6p-hydroxyalkyl pencil lanate sulfone with sulfur trioxide-pyridine complex in dimethylformamide at room temperature for 45-60 minutes.

The product, which is isolated as the pyridiniurn salt, is debenzylated with hydrogen and 5% palladium-on-calcium carbonate, as previously described.

When R, is an ester-forming residue readily hydrolyzable in vivo in a compound of formula I or 11 it is a group which is notionally derived from an alcohol of the formula R,-OH, such that the moiety COOR, in such a compound of formula I or 11 represents an ester grouping. Moreover, R, is of such a nature that the grouping COOR, is readily cleaved in vivo to liberate a free carboxy group (COOH). That isto say, R, is a group of the type that when a compound of formula I or 11, wherein R, is an ester-forming residue readily hydrolyzed in vivo, is exposed to mammalian blood or tissue, the compound of formula I or 11, wherein R, is hydrogen, is readily produced. The groups R, are known in the penicillin art. In most instances they improve the absorption characteristics of the pencillin compound. Additionally, R, should be of such a nature that it imparts pharmaceutically-acceptable properties to a compound of formula I or 11, and it liberates pharmaceutical ly-acceptable fragments when cleaved in vivo.

As indicated above, the groups R, are known and are readily identified by those skilled in the pencillin art, as taught in West German application 2,517,316. Typical groups for R, are 3-phthalidyl, 4-crotonolactony!, -y-butyrolacton-4-yi, alkanoyloxyalkyl and alkoxycarbonyloxyalkyl. However, preferred groups for R, are alkanoyloxymethyl having from 3to 6 carbon atoms, 1-(alkanoyloxy)ethyl having from 4to 7 carbon atoms, 30 1-methyl-l-(alkanoyloxy)ethyl having from 5to 8 carbon atoms, alkoxycarbonyloxymethyl having from 3 to 6 carbon atoms, 1-(alkoxycarbonyloxy)ethyl having from 4to 7 carbon atoms, 1-methyl-l (alkoxycarbonyloxy)ethyl having from 5 to 8 carbon atoms, 3-phthalidyl, 4crotonolactonyl and -y butyrolacton-4-yl.

-15 Compounds of the formula I or 11, wherein R, is an ester-forming residue readily hydrolyzable in vivo, can 35 be prepared directly from the compound of formula I or 11, wherein R, is hydrogen, by esterification. The specific method chosen will depend naturally upon the precise structure of the ester-forming residue, but an appropriate method will be readily selected by one skilled in the art. In the case where R, is selected from the group consisting of 3-phthalidyl, 4-crotonolactonyl, -y-butyrolacton-4-yi, alkanoyloxyalkyl and alkoxycar- bonyloxyalkyl they can be prepared by alkylation of the compound of formula I or[[, wherein R, is hydrogen, 40 with a 3-phthalidyl halide, a 4-crotonolactonyl halide, a -y-butyrolacton-4-yi halide, an alkanoyloxyalkyl halide or an alkoxycarbonyloxyalkyl halide. The term "halide" is intended to mean derivatives of chlorine, bromine and iodine. The reaction is conveniently carried out by dissolving a salt of the compound of formula I or 11, wherein R, is hydrogen, in a suitable polar organic solvent, such asNN-dimethylformamide, and then adding about one molar equivalent of the halide. When the reaction has proceeded essentially to completion, the product is isolated by standard techniques. It is often suff icient simply to dilute the reaction medium with an excess of water, and then extract the product into a water- immiscible organic solvent and then recover same by solvent evaporation. Salts of the starting material which are commonly used are alkali metal salts, such as sodium and potassium salt, and tertiary amine salts, such as triethylamine, N-ethylpiperidine, NN-dimethylaniline and N-methylmorpholine salts. The reaction is run at a temperature 50 in the range from about 0 to 50'C., and usually at about 0-25C. The length of time needed to reach completion varies according to a variety of factors, such as the concentration of the reactants and the reactivity of the reagents. Thus, when considering the halo compound, the iodide reacts faster than the bromide, which in turn reacts faster than the chloride. In fact, it is sometimes advantageous, when utilizing a chloro compound, to add up to one molar equivalent of an alkali metal iodide. This has the effect of speeding 55 up the reaction. With full regard for the foregoing factors, reaction times of from 1 to about 24 hours are commonly used.

The compounds of formula I and 11, wherein R, is hydrogen, are acidic and will form salts with basic agents. Such salts are considered to be within the scope of this invention. These salts can be prepared by standard techniques, such as contacting the acidic and basic componentc:, usually in a 1:1 molar ratio, in an 60 aqueous, non-aqueous or partially aqueous medium, as appropriate. They are then recovered by filtration, by precipitation with a non-solvent followed by filtration, by evaporation of the solvent, or in the case of aqueous solutions, by Iyophilization, as appropriate. Basic agents which are suitably employed in salt formation belong to both the organic and inorganic types, and they include ammonia, organic amines, alkali metal hydroxides, carbonates, bicarbonates, hydrides and alkoxides, as well as alkaline earth metal 65 7 GB 2 128 986 A 7 hydroxides, carbonates, hydrides and alkoxides. Representative examples of such bases are primary amines, such as n-propylamine, n-butylamine, aniline, cyclohexylamine, benzylamine and octylamine; secondary amines, such as diethylamine, morpholine, pyrrolicline and piperidine; tertiary amines, such as triethylamine, N-ethylpiperidine, N-methylmorpholine and 1,5-diazabicyclo [4.3.01non-5-ene; hydroxides such as sodium hydroxide, potassium hydroxide, ammonium hydroxide and barium hydroxide; alkoxides, 5 such as sodium ethoxide and potassium ethoxide; hydrides, such as calcium hydride and sodium hydride; carbonates, such as potassium carbonate and sodium carbonate; bicarbonates, such as sodium bicarbonate and potassium bicarbonate; and alkali metal salts of long-chain fatty acids, such as sodium 2 ethylhexanoate.

Preferred salts of the compounds of formula I and 11 wherein R, is hydrogen are the sodium, potassium 10 and triethylamine salts.

As indicated hereinbefore, the compounds of the formula I and 11, wherein R, is hydrogen or an ester-forming residue readily hydrolyzable in vivo, are potent inhibitors of microbial P-lactamases, and they increase the antibacterial effectiveness of p-lactam antibiotics (penicillins and cephalosporins) against many microorganisms, particularly those which produce a P-lactamase. The ability of the said compounds of the formula I or 11 increase the effectiveness of a P-lactam antibiotic can be appreciated by reference to experiments in which the MIC of a given antibiotic alone, and a compound of the formula I or 11 alone, are measured. These MIC's are the compared with the MIC values obtained with a combination of the given antibiotic and the compound of the formula I or 11. When the antibacterial potency of the combination is significantly greater than would have been predicted from the potencies of the individual compounds, this is 20 considered to constitute enhancement of activity. The MIC values of combinations are measured using the method described by Barry and Sabath in "Manual of Clinical Microbiology", edited by Lenette, Spaulding and Truant, 2nd edition, 1974, American Society for Microbiology.

The compounds of the formula I and 11, wherein R, is hydrogen or an esterforming residue readily hydrolyzable in vivo, enhance the antibacterial effectiveness of P-lactam antibiotics in vivo. That is, they 25 lower the amount of the antibiotic which is needed to protect mice against an otherwise lethal inoculum of certain P-lactamase producing bacteria.

The ability of the compounds of formula I and 11, wherein R, is hydrogen or an ester-forming residue readily hydrolyzable in vivo, to enhance the effectiveness of a p-lactam antibiotic against p-lactamase producing bacteria makes them valuable for co-administration with p- lactam antibiotics in the treatment of 30 bacterial infections in mammals, particularly man. In the treatment of a bacterial infection, the said compound of the formula I or 11 can be comingled with the P-lactam antibiotic, and the two agents thereby administered simultaneously. Alternatively, the said compound of the formula I or 11 can be administered as a separate agent during a course of treatment with a P-lactarn antibiotic. In some instances it will be advantageous to pre-dose the subject with the compound of the formula I or 11 before initiating treatment 35 with a P-lactam antibiotic.

When using a compound of formula I or 11 wherein R, is hydrogen or an ester group readily hydrolyzable in vivo to enhance the effectiveness of P-lactam antibiotic, a mixture of I or 11 with the P-lactarn antibiotic is administered preferably in formulation with standard pharmaceutical carriers or diluents. A pharmaceutical composition comprising a pharmaceutically-acceptable carrier, a P-lactam antibiotic and a compound of 40 formula I or 11 wherein R, is hydrogen or a readily hydrolyzable ester thereof will normally contain from about 5 to about 80 percent of the pharmaceutically acceptable carrier by weight.

When using the compounds of formula I or 11 wherein R, is hydrogen or an ester group readily hydrolyzable in vivo in combination with another P-lactam antibiotic, said compounds can be administered orally or parenterally, i.e. intramuscularly, subcutaneously or intra peritonea] ly. Although the prescribing physicial will ultimately decide the dosage to be used in a human subject, the ratio of the daily dosages of the compounds of formula I or 11 and the P-lactarn antibiotic will normally be in the range from about 1:3 to 3: 1.

Additionally, when using the compounds of formula I or 11 in combination with another P-lactam antibiotic, the daily oral dosage of each component will normally be in the range from about 10 to about 200 mg. per kilogram of body weight and the daily parenteral dosage of each component will normally be about 10 to 50 about 400 mg. per kilogram of body weight. These figures are illustrative only, however, and in some cases it may be necessary to use dosages outside these limits.

Typical P-lactam antibiotics with which the compounds of formula I or 11 and its esters readily hydrolyzable in vivo can be co-administed are:

6-(2-phenylacetamido)penicillanic acid, 6-(2phenoxyacetamido)penicillanic acid, 6-(2-phenylpropionamido)penicillanic acid, 6-(D-2-ami no-2-phenylaceta mido)penicil Ian ic acid, 6-(D-2-amino2-[4-hydroxyphenyl]acetamido)penicillanic acid, 60 6-(D-2-amino-2-[1,4cyclohexadienyllacetamido)penicillanic acid, 6-(1 -a mi nocyclohexanecarboxamiclo) pen icil la nic acid, 6-(2-carboxy-2phenylacetamido)penicillanic acid, 6-(2-ca rboxy-2-[3thienyllacetamido)peniciI Ian ic acid, 6-(D-2-[4-ethylpiperazin-2,3-dionel-carboxamidol-2-phenylacetamido)penicill anic acid, 65 6-(D-2-[4-hydroxy1,5-naphthyridine-3-carboxamidol-2-phenylacetamido)-penici llanic acid, 8 GB 2 128 986 A 8 6-(D-2-su Ifo-2-phenylaceta m ido) pen ici I I an ic acid, 6-(D-2-su Ifoam i no-2-p henyl acetam ido) pen ici I [an ic acid, 6-(D-2-[i m idazo I idi n-2-o ne-1 -ca rboxa m idol-2-phenyl acetam ido) pen ici Ilan ic acid, 6-(D-2-[3-methylsulfonylimidazolidin-2-one-l-carboxamido]-2phenylacetamido) penicillanic acid, 6-([hexahydro-1 H-azepin-1-yllmethyleneamino)penicillanic acid, acetoxymethyl 6-(2-phenylacetamido)penicillanate, acetoxymethyl 6-(D-2-amino-2-phenylacetamido)penicillanate, acetoxymethyl 6-(D-2-amino-2-[4-hydroxyphenyllacetamido)-penicillanate, pivaloyloxymethyl 6-(2-phenylacetamido) pen icil la nate, pivaloyloxymethyl 6-(D-2-amino-2-phenylacetamido)penicillanate, pivaloyloxymethyl 6-(D-2-amino-2-[4-hydroxyphenylacetamido)penicillanate, 1-(ethoxycarbonyloxy)ethyl 6-(2-phenylacetamido)penicillanate, 1 -(ethoxyca rbo nyloxy) ethyl 6-(D-2-amino-2phenylacetamido)penicillanate, 1-(ethoxycarbonyloxy)ethyl 6-(D-2-amino-2-[4- hydroxyphenyl)acetamido)penicillanate, 3-phthalidyl 6-(2-phenylacetam!do) pen icil la nate, 3-phthalidyl 6-(D-2-amino-2-phenylacetamido)penicillanate, 3-phthalidyl 6-(D-2-amino-2[4-hydroxyphenyllacetamido)-penicillanate, 6-(2-phenoxycarbonyl-2-phenylacetamido)penicillanic acid, 6-(2-tolyloxycarbonyl-2-phenylacetamido)penicillanic acid, 6-(2-[5-indanyloxycarbonyll-2-phenylacetamido)penicillanic acid, 6-(2-phenoxycarbonyl-2-[3-thienyllacetamidopenicillanic acid, 6-(2-tolyloxycarbonyl-2-[3-thienyllacetamido)penicillanic acid, 6-(2-[5-i nda nyl oxyca rbo nyl]-2-[3-th ienyl] aceta m ido) pen ici I [an ic acid, 6-(2,2-dimethyl-5-oxo-4-phenyl-l-imidazolidinyl)penicillanic acid, 7-(2-[2-thienyllacetamido)cephalosporanic acid, 7-(2-[l -tetrazo lyl I aceta m ido-3-(2-[5-m ethyl - 1,3,4-th iad i azolyl Ithio m ethyl)-3 desacetoxymethy1cephalosporanic acid, 7-(D-2-formyloxy-2-phenylacetamido)-3-(5-[1 -methyltetrazolyllthiomethyl)3- desacetoxymethy1cephalosporanic acid, 7-(D-2-amino-2-phenylacetamido)desacetoxycephalosporanic acid, 7-alpha-methoxy-7-(2-[2-thienyllacetamido)-3-carbamoyloxymethyl-3desacetoxy methylcephalosporanic acid, 7-(2-cyanoacetamido)cephalosporanic acid, 7-(D-2-hydroxy-2-phenylacetamido)-3-(5-[l-methyltetrazolyllthiomethyl)-3desacetoxymethy1cephalosporanic acid, 7-(D-2-amino-2-p-hydroxypehnylacetamido)desacetoxy-cephalosporanic acid, 7-(2-[4-pyridylthiolactamido)cephalosporanic acid, 7-(D-2-amino-2[1,4-cyclohexadienyllacetamido)cephalosporanic acid, 7-(D-2-amino-2-phenylacetamido)cephalosporanic acid, 7-[D-(-)-alpha-(4-ethyl-2,3-dioxo-l-piperazinecaboxamido)-alpha-(4hydroxyph enyl)acetamidol-3-[l- 40 methyl- 1,2-3,4-tetrazo I -5-yl)th io m ethyl 1-3-ce p hem-4-ca rboxyl ic acid, 7-(D-2-amino-2-phenylacetamido)-3-chloro-3-cephem-4-carboxylic acid, 7-[2-(2-amino-4-thiazolyl)-2-(methoximino)acetamidolcephalosporanic acid, [6R,7R-3-carbamoyloxymethyl-7(2Z)-2-methoxyimino (fur-2-yi[acetamido-ceph3-em-4-carboxylateI 7-([2-(2-aminothiazol-4-yl)acetamidol-3-[([1-2-dimethylaminoethyl)-1 Htetrazol-5-yllthio)methyllceph-3- 45 em-4-carboxylic acid, and a pharmaceutically acceptable salts thereof.

As will be appreciated by one skilled in the art, some of the above plactam compounds are effective when administered orally or parenterally, while others are effective only when administered by the parenteral route. When compounds of formula I wherein R, is hydrogen or an ester group readily hydrolyzable in vivo is to be used simultaneously (i.e. co-mingled) with a P-lactam antibiotic which is effective only on parenteral 50 administration, a combination formulation suitable for parenteral use will be required. When the compounds of formula I wherein R-, is hydrogen or an ester group is to be used simultaneously (co-mingled) with a P-lactam antibiotic which is effective orally or parenterally, combinations suitable for either oral or parenteral administration can be prepared. Additionally, it is possible to administer preparations of the compounds of formula I orally, while at the same time administering a further P-lactam antibiotic parenterally; and it is also possible to administer preparations of the compounds of formula I parenterally, while at the same time administering the further P-lactam antibiotic orally.

The following examples are provided solely for the purpose of further illustration. Nuclear magnetic resonance spectra (NMR) were measured at 60 MHz for solutions in deuterochloroform (CDC13). perdeutero dimethyl sulfoxide (DMSO-d6) or deuterium oxide (D20) or are noted otherwise, and peak positions are expressed in parts per million (ppm) downfield from teramethylsilane or sodium 2,2,-dimethyl-2 silapentane-5-sulfonate. The following abbreviations for peak shapes are used: s, singlet; d, doublet; t, triplet; q, quartet; m, multiplet.

J 9 GB 2 128 986 A 9 EXAMPLE 1

6P-Hydroxymethyl penicil Ian ic acid sulfone A. benzyl 6-bromo-6-hydroxymethylpenicillanate A solution of 44.9 g. of benzyl 6,6-dibromopenicillanate in 600 ml. of dry tetrahydrofuran was cooled to -78'C. and 56.4 ml. of t-butylmagnesium chloride was added dropwise with vigorous stirring under an inert atmosphere while maintaining the temperature at -60"C. After stirring 30 min. at -78'C. the solution was treated with gaseous formaldehyde in a stream of nitrogen until five molar equivalents had been added. The reaction was quenched at -78'C. by the addition of 5.7 ml. of acetic acid dropwise over a period of 25 min.

The reaction solution was allowed to warm to room temperature and was concentrated in vacuo. To the residue was added 200 ml. of water and 200 ml. of ethyl acetate. The organic layer was separated and the water layer extracted again with ethyl acetate. The organic phases were combined, washed successively with water (200 mi.), 5% aqeuous sodium bicarbonate (200 ml.) and brine (200 mi.) and dried over magnesium sulfate. Removal of the solvent under reduced pressure provides 38.2 g. of the desired product, epimeric at C-6.

B. benzyl 6p-hydroxymethylpenicillanate A solution containing 10 g. of benzyi 6-bromo-6- hydroxymethylpenicillanate,6.9 m]. tri-n-butyltin hydride and a trace of azobisisobutyronitrile in 200 mi. of benzene was refluxed for 5 hrs. under nitrogen. The reaction mixture was cooled and concentrated in vacuo. The residue was triturated with hexane and was chromatographed on silica gel, using toluene/ethyl acetate (2:1) as the eluent to give 7.5 g. of the product. 20 C. benzyl 6p-hydroxymethylpenicillanate sufflone m-Chloroperbenzoic acid (11.8 g.) was added to a solution of 7.5 9. of benzyi 6p- hydroxymethylpenicillanate in 600 mi. of methylene chloride cooled to O- WC. The solution was then allowed to warm to room temperature and was stirred for 5 hrs. The solvent was removed in vacuo and the residue partitioned between 200 mi. of water and 200 mi. of ethyl acetate. The pH of the mixture was adjusted to 7 by the addition of a saturated sodium bicarbonate solution, and sufficient sodium bisuifite was added to give a negative peroxide test (starch-iodide). The layers were separated, and the aqueous washed with ethyl acetate. The organic layer and washings were combined, washed successively with water, 5% sodium bicarbonate solution and brine and dried over magnesium sulfate. Removal of the solvent under reduced 30 pressure gave a foam, which on chromatographing on silica gel (chloroformethyl acetate 20:3) gave 3.5 g.

of the desired intermediate product.

D. calcium 6p-hydroxymethylpenicillanate sulfone To a 30 mi. solution of water-methanol 0: 1) was added 3.5 g. of 5% palladium on calcium carbonate and the catalyst prehydrogenated at 47 psi in a hydrogenation apparatus. To the resulting catalyst was added 3.5 g. of benzyi 60-hydroxymethylpenicillanate sulfone in 10 mi. of methanol and 20 mi. of tetrahydrofuran, and the mixture was shaken in a hydrogen atomsphere at 48 psi for 30 min. The catalyst was filtered through a filter aid and the filtrate concentrated in vacuo. The aqueous residue was extracted with ethyl acetate (2 x 100 mi.) and freeze dried to give 3.0 9. of the desired product as the calcium salt.

The NIVIR spectrum (CDC13 - free acid) showed absorption at 1.49 (s, 3H, 1.6 (s, 3H), 4.1 (m, 3H), 4.32 (s, 1 H) and 4.9 (cl, 1 H, J=41-iz) ppm.

EXAMPLE 2

6p-[l (S)Hydroxyethyllpenicillanic acid suffone A. benzyl 6p-bromo-6o--[l(S)hydroxyethyllpencillanate To a stirred solution of 9 g. of benzyl 6,6-dibromopencillanate in 200 ml. of dry toluene at -78'C. under argon was added dropwise 15 ml. of t-butyl lithium in pentane. The greenish-yellow solution was stirred 15 min. at -78'C, followed by the addition of 10 ml. of a 1 Msolution of anhydrous zinc chloride in tetrahydrofuran. The mixture was stirred 45 min. at -78'C., and 5 ml. of acetaldehyde was added while maintaining the temperature at -78'C. After one hour of additional stirring, the reaction was quenched with ml. of acetic acid in 50 ml. of ether at -78'C., and allowed to warm to room temperature. The toluene was removed in vacuo, and the residue partitioned between water and ethyl acetate. The organic phase was separated and the aqueous extracted (2 x 100 ml.) with ethyl acetate. The combined organic layers were washed with a 5% aqueous sodium bicarbonate solution followed by a saturated brine solution. They were 55 then dried over magnesium sulfate and concentrated to an oil, which on chromatographing on silica gel, using toluene-ethyl acetate as the eluent (10:2), gave 3.6 g. of benzyl 6oL-bromopenicillanate and 3.9 g. of the desired product. An analytical sample was purified by recrystallization from ether, m.p. 124-125'C.

The NMR spectrum (CDC13) showed absorption at 1.42 (s, 3H), 1.45 (d, 3H, J=6Hz), 1.67 (s, 3H), 2.55 (s, 1 H), 4.3 (q, 1H,J=6Hz), 4.6 (s, 1H), 5.3 (s, 2H), 5.5 (s, 1H) and 7.4 (s, 5H) at ppm.

B. benzyl 60-[1(S)Hydroxyethyllpenicillanate To 20 mi. of methanol-water(1:1) was added 1.1 g. of 5% palladium-on- calcium carbonate, and the resulting mixture was hydrogenated at 47 psi. for 20 min. To the resulting black slurry was added 1.1. g. of benzyl 6-p-bromo-69--[1 (S)hydroxyethyll pen icil la nate and the hydrogenation continued at 47 psi. for 30 min.65 GB 2 128 986 A The spent catalyst was removed by filtration and the filtrate concentrated under reduced pressure. The pH of the residual aqueous was adjusted to 8, and extracted with methylene chloride. The organic phase was separated, dried over magnesium sulfate and evaporated to an oil. The residual oil was then chromatographed on 150 g. of silica gel using chloroform-ethyl acetate (10: 1) as the eluent. Fractions 23 through 33 5 were combined and evaporated to dryness to give 148 mg. of the desired product.

The NMR spectrum (CIDC13) showed absorption at 1.35 (cl, 3H), 1.4 (s, 3H), 1.58 (s, 3H), 2.5 (m, 1 H), 3.45 (dd, 1 H, J=1 0, 4Hz), 4.2 (m, 1 H), 4. 38 (s, 1 H), 5.13 (s, 2H), 5.38 (d, 1 H, J=41-1z) and 7.33 (s, 51-1) ppm.

C. benzyl 6p-[l(S)Hydroxyethyllpencillanate suffione To a solution of 148 mg. of benzyl 60-[l (S) hyd roxyethyll pencil la nate in 20 mi. of methylene chloride at 0-5'C. was added portionwise 223 mg. of m-chloroperbenzoic acid. The resulting reaction mixture was allowed to stir at room temperature overnight. The undissolved solids were filtered and the filtrate evaporated under reduced pressure to dryness. The residue was partitioned between a 5% aqueous sodium bicarbonate solution and ethyl acetate, Sodium bisulfite was added to the vigorously stirred mixture until a negative peroxide test (starch iodide) was obtained. The organic layer was subsequently separated and the 15 aqueous layer extracted with additional ethyl acetate. The combined organic layers were successively backwashed with a saturated sodium bicarbonte solution and brine solution, and were then dried over magnesium sulfate. Removal of the solvent in vacuo gave 160 mg. of the product as an oil.

The NIVIR spectrum (CD13) showed absorption at 1.27 (s, 3H), 1.35 (d, 3H, J=6Hz), 1.5 (s, 3H), 3.2 (m, 1 H), 3.85 (dd, 1H,J=11, 5Hz), 4.53 (s, 1H), 4.77 (m, 1H), 4.77 (d, 1H,J=5hz), 5.28 (ABq, 2H,J=12Hz) and 7.43 (s, 5H) 20 ppm- D. 6P-[1(S)hydroxyethyllpenicillanic acid sufflone A suspension of 160 mg. of 5% palladium-on-calcium carbonate in 20 m]. of methanol-water (1:1) was prehydrogenated at 47 psi. for 20 min. To the resulting suspension was added 160 mg. of benzyi 60-[1 (S)-hydroxyethyllpenicillanate suifone and the hydrogenation continued at 51 psi. for one hour. The spent catalyst was filtered and the pH of the filtrate adjusted to 8. Afterthe aqueous was extracted with ethyl acetate, the pH was adjusted to 1.8 and fresh ethyl acetate added. The ethyl acetate layer was separated, dried over magnesium sulfate and concentrated to an oil, 90 mg. The oil subsequently crystallized to give a white solid, m.p. 160-161.5'C. dec.

The NMR spectrum (CDC13 - DiVISO - D6) showed absorption at 1.2 (d, 3H, J=61-1z), 1.42 (s, 3H), 1.52 (s, 31-1), 3.80 (dd, 1 H, J=10, 5Hz), 4.28 (s, 1 H), 4.5 (m, 1 H) and 5.20 (d, 1H, J=51-1z) ppm.

EXAMPLE 3

6p-[l (R)Hydroxyethyll pen icil lanic acid sulfone A. benzyl6p-bromo-6o--[l(R)hydroxyethyllpencillanate To 50 ml. of toluene cooled to -78'C. was added slowly 70 ml. of diethyl zinc, also cooled to -78'C.

Subsequently, 45 g. of benzyl 6,6-dibromopenicillanate in 250 ml. of toluene was added to the reaction mixture over a period of 45 min. After one hour of stirring in the cold 17 ml. of acetaldehyde was added to the reaction mixture, and the stirring continued for one hour. The reaction was quenched by the addition Of 11.5 40 ml. of acetic acid in 100 mi. of diethyl ether. The cooling bath was removed and the reaction allowed to warm to room temperature. An equal volume of water and ethyl acetate was added to the reaction mixture and allowed to stirfor 5 min. The organic phase was subsequently separated and washed successively with water (3 X 75 ml.), a saturated sodium bicarbonate solution (3 X 75 mi.) and a saturate brine solution (1 X 100 ml.). The organic layer was dried over magnesium sulfate and evaporated in vacuo to an oill which was 45 chromatographed on 500 g. of silica gel using chloroform-ethyl acetate (10:1) as the eluent. Fractions 13 through 29 were combined and evaporated to dryness to give 20 g. of the crude intermediate product which was recrystallized from diethyl ether-hexaneto give 12.7 g., m.p. 109-1 10'C. The isolated material also contained benzyl 6p-bromo-6-a-[l(S)hydroxyethyl]penicillanate.

B. benzy] 6P-[1(R)hydroxyethyl]penicillanate A solution of 1.0 g. of benzyl 6p-bromo-6ot-[1 (R)hydroxyethyllpenicillanate and 1.4 mi. of tri-n-butyltin hydride in 35 mi. of benzene, under a nitrogen atomsphere was refluxed for 40 min. The reaction mixture was then cooled to room temperature and the solvent removed under reduced pressure. The residue was triturated repeatedly with hexane. The residue was chromatographed on 100 9. of silica gel using chloroform-ethyl acetate (20:1) as the eluent. Fractions 82 through 109 were combined and evaporated to give 750 mi. of the desired product.

The NMR spectrum (CDC13) showed absorption at 1.18 (d, 3H, J=61-1z), 1.38 (s, 3H), 1.62 (s, 3H), 2.6 (n, 1 H, 3.45 (dd, 1 H, J=9,41-1z), 4.2 (m, 1 H), 4.43 (s, 1 H), 5.16 (s, 2H), 5.33 (d, 1 H, J=4Hz) and 7.33 (s, 5H) ppm.

C. benzyl 60-[1(R)hydroxethyllpenicillanate sufflone A mixture of 335 mg. of benzyi 6p-[1 (R)hydroxyethyllpenicillanate and 507 mg. of m-chloroperbenzoic acid in 50 mi. of methylene chloride is allowed to stir overnight at room temperature. The solids are filtered and the solvent removed from the filtrate. The residue is partitioned between 50 mi. of water and 50 m[. of ethyl acetate. Sodium bisulfite is added in portions to the stirred mixture until all the peroixde is destroyed as 65 4 11 GB 2 128 986 A 11 evidence by a negative starch-iodide test. The oganic phase is separated, dried over magnesium sulfate and the solvent removed in vacuo. The residue is utilized in the next step without further purification.

D. 60-[1(R(hydroxyethyllpenicillanic acid sufflone A suspension of 1.78 g. of 5% palladium-on-calcium carbonate in 40 mi. of methanol-water (1: 1J is 5 hydrogenated for 20 min. at 50 psi. Benzyl 6p[(R)hydroxyethyllpenicillanate suifone (1.67 g.) is added to the resulting suspension and the hydrogenation continued at 50 psi for one hour. The methanol is removed under reduced pressure, and the aqueous residue extracted with ethyl acetate. The aqueous layer is acidified to pH 2 and extracted with ethyl acetate. The organic phase is washed with a saturated brine solution, dried over magnesium sulfate and concentrated to a white solid, 1.0 g., m.p. 182-1183'C. dec.

The NIVIR spectrum (DIVISO - D6) showed absorption at 1.15 (cl, 3H,J=6Hz), 1.37 (s, 3H,) 1.47 (s, 31-1), 3.87 (dd, 1H,J=10,5Hz),4.28 (s, 1WA.5 (m, 11-1), 5.11 (cl, 1H,J=5Hz) and 5.5 (mAH).

EXAMPLE 4

6P-[1(S)Hydroxyethyllpenicillanic acid suifone A. benzyl 6p-bromo-6oL[1(S)hydroxyethyljpenicillanate sufflone To 500 mi. of methylene chloride, maintained at 5'C. under a nitrogen atmosphere, was added 14.7 g. of benzyi 6p-bromo-6oL-[1 (S)hydroxyethyllpenicillanate 14.7 g. of benzyi 6p-bromo-60t[1 (S)hydroxyethyllpenicillanate (Example 2A) and 17.8 g. of m- chloroperbenzoic acid, and the resulting reaction mixture allowed to stir overnight. An additional 200 mg. of the per acid is added and stirring continued for an additional 2.5 hrs. The reaction was filered and the filtrate concentrated to a white solid. An equal volume of water-ethyl acetate was added to the residue and the pH adjusted to 7.4 with a saturated sodium bicarbonate solution. The organic phase was separated, added to fresh water and the pH adjusted to 8.2 with a saturated sodium bicarbonate solution. The ethyl acetate layer was backwashed with a saturated sodium bicarbonate solution (3 X 400 mi.) and then with a brine solution. The oganic phase was separated, 25 dried over magnesium sulfate and evaporated to an oil, 18.2 g.

The NIVIR spectrum (CIDC13) showed absorption at 1.28 (s, 31-1), 1.43 (d, 3H, J=61-1z), 1.55 (s, 31-1), 4.2 (q, 1 H, J=61-1z), 4.57 (s, 1 H), 4.85 (s, 1 H), 5.23 (ABq, 2H, J=121-1z) and 7.38 (s, 51-1) ppm.

B.benzyl6p-[1(S)hydroxyethyllpenicillanatesulfone To a solution of 740 mg. of benzyi 6p-bromo-6a.- [1(S)hydroxyethyllpenicillanatesuifone in 10 m[. of benzene under a nitrogen atomsphere was added.52 mi. of tri-n-butyitin hydride, and the resulting mixture heated under reflux for 3 hrs. The benzene was removed in vacuo and the residue triturated with hexane. The hexane was decanted and the residue used in the next step without further purification.

C. 6p-[l(S)hydroxyethyljpenicillanic acid sulfone One gram of 5% pall adiu m-on-calcium carbonate in 20 ml. of water was prereduced with hydrogen at 50 psi. To the resulting slurry was added the crude benzyl 6p-[l (S)hydroxyethyllpenicillanate sulfone in 20 mi.

of methanol from Example 4B, and hydrogenation continued for one hour at 50 psi. An additional 500 mg. of catalystwas added and the reaction continued for 45 min. The spent catalyst was filtered and the filtrate 40 extracted (2 x 50 mi.) with ethyl acetate. The aqueous was over laid with fresh ethyl acetate on the pH adjusted to 1.5. The organic phase was separated and the aqueous extracted with ethyl acetate (7 X 100 ml.).

The ethyl acetate acid extracts were combined, washed with a brined solution, dried over magnesium sulfate and concentrated in vacuo to dryness, 230 mg.

The NIVIR spectrum was identical to the product prepared in Example 2D.

EXAMPLE 5

6p-[1 (R)Hydroxyethyllpenicillanic acid suifone A. benzyl 6p-bromo69--[1 (R)hydroxyethyllpenicillanate sufflone To a solution of 2.9 g. of benzyl 6p-bromo-6a-[ 1 (R)-hydroxyethyll pen icil lanate (Example 3A) in 100 mi. of methylene chloride cooled to 0-5'C. was added 3.6 g. of mchloroperbenzoic acid, and the resulting reaction mixture allowed to stir at room temperature overnight. The solvent was removed under reduced pressure and the residue dissolved in an equal volume of water-ethyl acetate. The pH of the mixture was adjusted to 7.4 with a saturated sodium bicarbonate solution and the organic layer separate. The organic phase was back-washed with a saturated brine solution, dried over magnesium sulfate and concentrated to an oil which 55 crystallized, 4.0 g. The NIVIR spectrum (CIDC13) showed absorption at 1.25 (s, 3H), 1.28 (cl,

3H, J=61-1z), 1.5 (s, 3H), 2.9 (m, 1 H), 3.7 (dd, 1 H, J=110, 5Hz), 4.43 (s, 1 H), 4.6 (m, 1 H), 4.57 (cl, 1 H, J=51-1z), 5.17 (ABq, 2H, J=121-1z) and 7.32 (s, 51-1) 13pm.

B. benzyl 6P-[1(R(hydroxyethyllpenicillanate sufflone A mixture of 3.0 9. of benzyi 6p-bromo-6u-[1 (R)- hydroxyethyllpenicillanate suifone and 2.9 m I. of tri-n-butyltin hydride in 100 mi. of benzene was refluxed under a nitrogen atmosphere for 30 min. The solvent was removed in vacuo and the residue extracted several times with hexane. The residual material 12 GB 2 128 986 A 12 was chromatographed on 250 g. of silica gel to give 1.67 g. of the desired product, which was used in the next step.

C. 6P-[1(R)hydroxyethyllpenicillanic acid sufflone One and seven-tenths grams of 5% palladium-on-calcium carbonate in 40 m]. of 50% methanol-water was 5 pre-reduced at 50 psi for 20 min. To the resulting suspension was added 1. 67 g. of benzy] 6p [(R)hydroxyethyll-pencillanate sulfone, and the hydrogenation continued for one hour. The catalyst was filtered and the methanol removed in vacuo from the filtrate. The aqueous residue was extracted with water, followed by the adjustment of the aqueous to 2.0. The acidified aqueous was extracted several times with ethyl acetate, and the combined extracts washed with a saturated brine solution and dried over magnesium 10 sulfate. Removal of the solvent gave 1.0 9. of the product, which was indistinguishable in all respects from that prepared in Example 3D.

EXAMPLE 6

Starting with benzy] 6,6-dibromopenicillanate and the appropriate aldehyde and employing the procedures of Example 2, the following 6p-[1 (S)hydroxyalkyllpenicillanic acid sulfones are prepared: 6p[l (S)hydroxypropyllpenicillanic acid suifone; 6p-[1 (S)hydroxy-nbutyllpenicillanic acid suifone; 6p[l (S)hydroxy-3methylpropyllpenicillanic acid sulfone;6p[l (S) hyd roxy-2-m ethyl propyl 1 penici 11 anic acid sulfone;6p[l (S)hydroxy-n-pentyllpenicillanic acid suifone; 60[1 (S) hyd roxy-2-m ethyl-n-butyl]pencil la nic acid sulfone;6p[l(S)hydroxy-3-methyi-n-butyllpenicillanic acid sulfone; and 6P[1(S)hydroxy-2,2dimethylpropyllpenicillanic acid sulfone.

EXAMPLE 7

Employing the procedures of Example 3, and starting with the requisite aldehyde and benzyl 6,6-dibromopencillanate, the following compounds are prepared:6P-[1 (R)hydroxypropyllpenicillanic acid 25 sulfone;60-[11 (R)hydroxy-2-methylpropyllpenicillanic acid sulfone;6p-[11 (R)hydroxy-n-pentyllpenicillanic acid suifone;6p-[1 (R)hydroxy-2-methyi-n-butyllpenicillanic acid sulfone; 6p- [11 (R)hydroxy-3-methyi-n butyllpenicillanic acid sulfone; and 6p-[1 (R)hydroxy-2,2-dimethylpropyll pen icillanic acid sulfone.

EXAMPLE 8 Starting with the appropriate benzyi 6-bromo-6[1-hydroxy alkyllpenicillanate and employing the procedures of Example 4/5, the following 6p-compounds are prepared:

R4 CH3CH2- (S) CH3CH2- (R) C1-13(CHA2- (R) 45 C1-13(CHA3- (S) C1-13(C1-12)3 (R) CH3CH(CH3)- (S) CH3CH(C1-13)CH2- (S) (CH3)3C- (R) 50 (C1-13)3C_ (S) ? H H H 0 2 CH R -CH 5 CH 4 3 0 1 C0 2 H EXAMPLE 9

6p-[l(S)Hydroxybenzyllpenicillanic acid sulfone and 6P41 (R)Hydroxybenzyllpenicillanic acid sulfone A. benzyl 6u--bromo-6P-[1(R) and (S)hydroxybenzyllpenicillanate To a solution of 9.0 g. of benzyl 6,6-dibromopenicillanate in 200 ml. of dry toluene cooled to -78'C. and maintained under an argon atmosphere was added 14 mi. of t-butyl lithium cooled to -67'C. After stirring in 60 the cold for 45 min. 2 mi. of benzaldehyde was added and the reaction mixture allowed to stir for one hour. A solution of 1.2 ml. of acetic acid in 50 mi. of diethyl ethyl was added over a period of 10 min. and the mixture stirred at -78'C. for 30 min. Water (100 ml.) and diethyl ethyl (100 ml.) were added and the mixture stirred at room temperature for 30 min. The organic layer was separated and the aqueous washed with ether. The organic layer and the ether washings were combined and washed successively with water (1 x 50 ml.), a A 1 13 GB 2 128 986 A 13 saturated aqueous sodium bicarbonate solution (2 X 50 ml.) and a saturated brine solution. The organic phase was dried over magnesium sulfate and concentrated to give 10.3 g. of an oil.

The residual material was chromatographed on 450 g. of silica gel using chloroform-ethyl acet ate (20: 1) as the eluent. Fractions 71 thru 101 were combined and concentrated to give 1.97 g. of the product as a 5 semisolid.

B. benzyl 6P-[l(R)hydroxybenzyllpenicillanate and benzyl 6p[l(S)hydroxybenzyllpenicillanate A solution of 1.9 g. of benzyl 6ot-bromo-6p-[1 (R) and (S)hydroxybenzyl] pen icil lanate and 1.1 ml. of tri-n-butyltin hydricle in 30 mi. of dry benzene under a nitrogen atmosphere was refluxed 3.5 hrs. An additional 1.0 mi. of hydride was added and the refluxing continued overnight. The benzene was removed in 10 vacuo and the residue was slurried with hexane. The hexane was decanted and the 850 mg. of residual oil chromatographed on 100 g. of silica gel using chloform-ethyl acetate (20:3) as the eluent. Fractions 20 thru 34 were combined and the solvent removed to give 495 mg. of benzyl 6p-[l(Rjhydroxybenzyllpenicillanate.

The NIVIR specturm (CDC13) showed absorption at 1.42 (s, 3H), 1.67 (s, 3H), 3.2 (m 1 HJ 3.9 (dd, 1 H, J=4, 1OHz),4.42 (s, 1H), 5.2 (s, 2H), 5.2 (m, 1H), 5.4 (d, 1H,J=4Hz) and 7.35 (m, 10H) ppm.

Fractions 35 thru 58 were combined and concentrated in vacuo to give 380 mg. of benzyl6p[1(S)hydroxybenzyllpenicillanate.

The NIVIR spectrum (CDC13) showed absorption at 1.33 (s, 3H), 1.67 (s, 3H), 3.4 (m, 1H), 3.85 (dd, 1H,J=4, 1OHz),4.42 (s, 1h, 5.10 (d, 1H,J=4Hz), 5.10 (s, 2H), 5.10 (m, 1H) and 7.35 (m, 10H) ppm.

C. benzyl 6p-[l(R)hydroxybenzyllpenicillanate sulfone To a solution of 490 mg. of benzyl 6p-[(R)hydroxybenzyl]penicillanate in 50 ml. of methylene chloride cooled to -5'C. was added 1.35 g. of m- chloroperbenzoic acid, and the resulting reaction mixture allowed to stir overnight. The solvent is removed in vacuo and the residue treated with an equal volume of ethyl acetate and water. The pH of the mixture is adjusted to 7.2 with a saturated sodium bicarbonate solution and sufficient sodium bisulfite was added to decompose the excess per acid (negative starch iodide test). The organic phase was separated and washed successively with water at a pH of 8.2, a saturated sodium bicarbonate solution and a brine solution. The organic layer was separated, dried over magnesium sulfate and concentrated to 395 mg. of a white solid. The product was chromatographed on 100 g. of silica gel using toluene-ethyl acetate (10:12) as the eluent. Fractions 18thru 27 were combined and concentrated to give 148 30 mg. of the product as an oil.

The NIVIR spectrum (CDC13) showed absorption at 1.22 (s, 3H), 1.5 (s, 3H), 2.6 (m, 1 H), 4.07 (dd, 1 H, J=1 0, 5Hz), 4.47 (s, 1 H), 4.67 (cl, 1 H, J=5Hz), 5.2 (ABq, 2H), 5.63 (cl, 1 H, J=1101Hiz) and 7.37 (m lOH) ppm.

W. benzyl 6P-[1(S)hydroxybenzyllpenicillanate sufflone The procedure of Example 9C was repeated starting with benzyi 6p-[11 (S)hydroxybenzyilpenicilianate to give the desired product.

The NIVIR spectrum (CDC13) showed absorption at 1.19 (s, 3H), 1.5 (s, 3H), 2.8 (m 1 H), 4.20 (dd, 1 H, J= 10, 5Hz), 4.38 (d, 1 H, J=5Hz), 4.43 (s, 1 H), 5.20 (ABq, 2H), 5.77 (d, 1 H, J=1 0Hz) and 7.37 (m, 1 OH) ppm.

D. 60-[l(R)hydroxybenzyljpenicillanic acid sulfone To a suspension of 148 mg. of 5% palladium on calcium carbonate in 20 ml. of water-methanol (1:1) which has been prehydrogenated at 47.5 psi for 20 min. was added 140 mg. of benzyl 6p [1 (R) hydroxybenzyll pen icil lante sulfone and the hydrogenation continued at an initial pressure of 47 psi for 40 min. An additional 140 mg. of catalyst was added and the reduction continued for 30 min. A final 140 mg- 45 of catalyst was then added and the reduction continued for an additional 30 min. The spent catalyst was filtered and the filtrate extracted with ethyl acetate. The aqueous layer was separated, the pH adjusted to 1.5 and fresh ethyl acetate added. The ethyl acetate extract was back-washed with a brine solution and dried over magnesium sulfate. Removal of the solvent in vacuo gave 90 mg. of the product as an oil.

The NIVIR spectrum (CDC13) showed absorption at 1.50 (s, 3H), 1.67 (s, 3H), 4.1 (dd, 1 H, J= 10, 5Hz), 4.45 (s, 50 1 H), 4.78 (d, 1 H, J=5Hz), 5.7 (d, 1 H, J = 1 OHz) and 7.4 (m, 5H).

W. 6p-[1 (S)hydroxybenzyllpenicillanic acid sufflone To a suspension of 170 mg. of prereduced 5% palladium-on-calclum carbonate in 20 mi. of water-methanoi (M) was added 170 mg. of benzyi 6p-[1(S)hydroxybenzyil-penicillanate suifone and the hydrogenation continued at 47 psi for 40 min. An additional 340 mg. of catalyst was added and the reduction was continued for 3 hrs. The catalyst was filtered, washed with tetra hyd rof u ra n- water (1: 1) and the combined filtrate and washings concentrated. The residual water was extracted with ethyl acetate followed by acidification of the aqueous layer to pH 1.5 and subsequent extraction with fresh ethyl acetate. The organic phase was backwashed with a brine solution and dried over magnesium sulfate. Removal of the solvent in vacuo gave 60 100 mg. of the product, m.p. 164165'C. dec.

The NIVIR (CDC13) showed absorption at 1.40 (s,3H), 1.55 (s,3H),4.0 (dd, 1H,J=5, 10Hz),4.4(d, 1H,J=5Hz), 4.4 (s, 1H), 5.7 (cl, 1H,J=10Hz), and 7.4 (m, 5H) pipm.

14 GB 2 128 986 A 14 EXAMPLE 10

6p-[1 (S)Hydroxy-2-phenethyllpenicillanic acid sulfone and 6p-[1 (R)Hydroxy-2-phenethyll pen icillanic acid sulfone A. benzyl 6ot-bromo-6p-[1(R)Hydroxy-2-phenethylpenicillanate and benzyl 6p-bromo-6(x-[1(S)hydroxy-2- 5 phenethyllpenicillanate To a solution of 9.0 g. of benzyl 6,6-dibromopenicillanate in 200 mi. of toluene cooled to -78'C. and maintained under an argon atmosphere was added 9.2 mi. of a 2.5M solution of t-butyl lithuim, and the resulting reaction mixture allowed to stir for 40 min. Subsequently, 2.34 mi. of phenyl acetal dehyde was 1() added. After stirring for one hour, 1.2 mi. of acetic acid in 25 m]. of diethyl etherwas added and the stirring 10 continued at -78'C. for 30 min. The reaction mixture was allowed to warm to room temperature afterwhich an equal volume of water was added. The organic layerwas separated and saved, and the aqueous was extracted with ethyl acetate. The organic layer and extracts were combined, washed successively with water, a saturated sodium bicarbonate solution and a brine solution and dried over magnesium sulfate. The oil residue, 11.0 g., which remained after the solvent was removed in vacuo was chromatographed on 500 g. of 15 silica gel using cloroform-ethyl acetate (20:0.2) as the eluent.

Fractions 150 thru 154 were combined and concentrated to give 670 mg. of benzy] 6a-bromo-6p[1(R)hydroxy-2-phenethyllpenicillanate.

The NMR spectrum (CDC13) showed absorption at 1.35 (s, 1 H), 1.53 (s, 1 H), 2.85 (m, 3H), 4.23 (m, 1 H), 4.41 (s, 1H),5.13 (s, 2H), 5.57 (s, 11-1) and 7.33 (m, lOH) ppm.

Fractions 155 thru 195 were combined and concentrated to give 4.84 g. of benzyl 6p-bromo-6ot[1(S)hydroxy-2-phenethyllpenicillanate.

The NMR spectrum (CDC13) showed absorption at 1.35 (s, 3H), 1.60 (s, 3H), 2.85 (m, 3H), 4.23 (m, 1 H), 4.41 (s, 1 H), 5.08 (s, 2H), 5.42 (s, 1 H) and 7.33 (m, 1 OH) ppm.

B. benzyl 6P-[1(R)hydroxy-2-phenethyllpenicillanate and benzyl 6P[1(S)hydroxy-2-phenethyllpenicillanate A benzene solution (80 mi.) containing 5.51 g. of benzyl 6ot-bromo-6p-[1 (R)hydroxy-2phenethyll pencil lanate and benzyl 60-brom o-6a-[(S) hyd roxy-2-phen ethyl] pen ici 11 anate, as isolated in Example 10A, was treated with 3.2 ml. of tri-n-butyltin hydride, and the reaction heated to reflux under a nitrogen atmosphere for 4 hrs. The solvent was removed under reduced pressure and the residue washed several times with hexane. The residue, 4.2 g., was chromatographed on 500 g. of silica gel using chloroform-ethyl acetate (20:3) as the eluent.

Fraction 50 thru 61 were combined and concentrated to give 596 mg. of benzyl 6P-[1(S)hydroxy-2phenethyllpenicillanate.

The NMR spectrum (CDC13) showed absorption at 1.35 (s, 3H), 1.69 (s, 3H), 2.8 (m, 2H), 3.1 (m, 1 H), 3.55 (dd, 35 1 H, J=4, 1 0Hz), 4.23 (m, 1 H), 4.40 (s, 1 H), 5.15 (s, 1 H), 5.35 (d, 1 H, J=4Hz), 7.22 (s, 51-1) and 7.3 (s, 51-1) ppm.

Fractions 65 thru 75 were combined and concentrated to give 1.5 g. of benzyl 6P-[1(R)hydroxy-2 phenethyllpenicillanate.

The NMR spectrum (CDC13) showed absorption at 1.35 (s, 3H), 1.6 (s, 3h), 2.78 (m, 2H), 2.9 (m, 1 H), 3.43 (dd, 1H, J=5.91-1z), 4.30 (m, 1H), 4.40 (s, 1H), 5.12 (s, 1H), 5.22 (cl, 1H, J=5Hz),7.119 (s, 5H) and 7.3 (s, 51-1) ppm. 40 C. benzyl 6P-[1(Sj-hydroxy-2-phenethyllpenicillanate sufflone To a cooled (O-WC.) solution of 300 mg. of benzyl 6p-[1 (S) hyd roxy-2-p henethyl 1 pencil 1 ante in 50 mi. of methylene chloride was added 630 mg. of m-chloroperbenzoic acid, and the resulting reaction mixture allowed to stir overnight. The solvent was removed in vacuo and the residue treated with an equal volume of 45 water and ethyl acetate. The pH of the mixture was adjusted to 7.2 with a saturated sodium bicarbonate solution, and sufficient sodium bisuifite was added to give a negative starch-iodide test. The organic phase was separated treated with an equal volume of water and the pH adjusted to 8.2 as above. The organic phase was separated, washed with a brine solution and dried over magnesium sulfate. Removal of the solvent gave the product as an oil, 320 mg.

The NMR spectrum (C13C13) showed absorption at 1.22 (s, 1 H), 1.5 (s, 1 H), 2.8 (m, 2H), 3.8 (dd, 1 H, J=5, 10Hz), 4.42 (s, 1 H), 4.6 (cl, 1 H, J=5Hz), 4.75 (m, 1 H) 5.18 (ABq, 2H), 7.2 (s, 51-1) and 7.3 (s, 5H) ppm.

C. benzyl 6P-[1(R)hydroxy-2-phenethyllpencillanate sulfone Using the procedure of Example 1 OC, and starting with 700 mg. of benzyl 6p-[1 (R)hydroxy-2-phenethyll- 55 penicillanate and 850 mg. of m-chloroperbenzoic acid there was obtained 610 mg. of the desired product as an oil.

The NMR spectrum (CDC13) showed absorption at 1.25 (s, 1 H), 1.52 (s, 1 H), (m, 2H), 3.7 (dd, 1H, J=5, 10Hz), 4.42 (s, 1 H), 4.55 (d, 1 H, J=5Hz), 4.80 (m, 1 H), 5.18 (ABq, 2H), 7.22 (s, 51-1) and 7.3 (s, 51-1) ppm.

D. 6p-[l(R)hydroxy-2-phenethyllpenicillanic acid sultone sodium salt To a suspension of 600 mg. of 5% palladium-on-calcium carbonate, prehydrogenated at 47 psi for 20 min., in 20 ml. of water-methanol (1:1) was added 600 mg. of benzyl 6p-[l (R)hydroxy-2-phenethyllpencillanate sulfone. After continuing the hydrogenation for 35 min. at 48 psi, an additional 600 mg. of catalyst was added and the hydrogenation at 48 psi continued for 10 min. The spent catalyst was filtered and washed with 65 GB 2 128 986 A 15 water-methanol (1:1). The filtrate and wash ings were combined and the methanol removed under reduced pressure. The residual aqueous (pH 8.0) was extracted with ethyl acetate and the layers separated. The aqueous layer was acidified to pH 1.8 and extracted with ethyl acetate. The organic phase was separated, backwashed with a brine solution and dried over magnesium sulfate. Removal of the solvent in vacuo gave 5 390 mg. of the product as a white solid.

The free acid was dissolved in ethyl acetate containing a small amount of diethyl ether. To this solution was added 177 mg. of sodium 2ethylhexanoate, and the solution allowed to stir for one hour. The precipitated solid sodium salt of the product was filtered and dried, 250 mg., m.p. 205-208'C. dec.

The NIVIR spectrum (D20) showed absorption at 1.42 (s, 3H), 1.65 (s, 3H), 2.9 (m, 2H), 4.0 (dd, 1 H, J=5, lo 1OHz),4.3 (s, 1H),4.9 (m, 1H),5.0 (d, 1H,J=5Hz) and 7.3 (s, 5H) ppm.

W. 60-[1(S)hydroxy-2-phenethyllpenicillanic acid sufflone To a suspension of 320 mg. of prereduced 5% palladium-on-calcium carbonate in 20 mi. of water-methanol (M) was added 320 mg. of benzyi 6p-[11 (S)hyd roxy-2-phenethyll pen icillanate suifone, and the resulting mixture was shaken in a hydrogen atmosphere at an initial pressure of 47 psi for 30 min. The catalyst was 1 filtered, washed with water-methanol, and the washings and filtrate were combined. The aqueous residue remaining after the methanol was removed in vacuo was extracted with ethyl acetate and was then extracted with fresh ethyl acetate. The extract was backwashed with a brine solution, dried over magnesium sulfate and evaporated to an oil, 80 mg, which solidified, m.p. 80-85'C. dec.

The NIVIR spectrum (C13C13) showed absorption at 1.42 (s, 1 H), 1.65 (s, 1 H), 2.9 (m, 2H), 4.0 (dd, 1 H, J=5, 20 10Hz), 4.3 (s, 1 H), 4.8 (m, 1 H), 4.85 (cl, 1 H, J=5Hz) and 7.3 (s, 5H) ppm.

EXAMPLE-11

6P-[1 (R)Hydroxy-3-phenylpropyllpenicillanic acid sulfone and 6P-[1 (S) Hydroxy-3-phenyl pro pyl] pen icil lanic acid sulfone A. benzyl 6a-bromo-6P-[1(R) and (S)hydroxy-3-phenylpropyllpenicillanate Benzyl 6,6-dibromopenicillanate (4.5 g.) was dissolved in 100 ml. of dry toluene and the resulting solution cooled to -70'C. To the cooled solution was added 7.3 mi. of t-butyl lithium. After stirring in the cold for 20 min. 1.3 mi. of hydrocinnarnaldehyde was added and stirring continued for 20 min. Acetic acid (0.57 ml.) was added and the reaction mixture allowed to warm to room temperature. The toluene was removed in vacuo and an equal volume of chloroform and water were added. The organic phase was separated, backwashed with a brine solution and dried over magnesium sulfate. Removal of the solvent gave 5.3 g. of the product as an oil. The product was purified by chromatographing on silica gel using chloro-form ethyl acetate (20: 1) as the eluent.

Fractions 88 thru 155 were combined and the solvent removed in vacuo to give 3.2 g. of the product.

Anal. Calc'd for C24H2004NSBr: C, 57.2; H, 5.2; N, 2.8. Found: C, 56.5; H, 5.2; N, 2.9.

B. benzyl 6P-[1(R)hydroxy-3-phenylpropyllpenicillanate andbenzyl 6P[1(S)hydroxy-340 phenylpropyllpencillanate A solution of 1.5 g. of benzyi 6a,-bromo-6p-[1 (R) and (S) hyd roxy-3- phenyl pro pyll pen icillanate and 1.72 ml.

of tri-n-butyltin hydride in 100 m[. of benzene was refluxed under a nitrogen atmosphere for 2 hrs. and 40 min. The solvent was removed in vacuo and the residue (3.7 g.) chromatographed on 150 g. of silica gel using chloroform-ethyl acetate (20: 1) as the eluent.

Fractions 63-80 were combined and the solvent removed to give 244 mg. of benzyl 6P-[1(S)hydroxy-3- 45 phenylpropyllpenicillanate as an oil.

The NIVIR spectrum (CDC13) showed absorption at 1.40 (s, 3H), 1.50 (s, 311), 1,8 (m, 211), 2.8 (m, 3H), 3.59 (dd, 1 H, J=4, 1 0Hz), 4.1 (m, 1 H), 4.43 (s, 1 H), 5.20 (s, 21-1), 5.43 (d, 1 H, J=4Hz), 7.25 (s, 51-1) and 7.4 (s, 5H) ppm.

Fractions 114-133 were combined and the solvent evaporated to give 369 mg. of benzyl 6P-[1(R)hydroxy-3- phenylpropyllpenicillanate as an oil.

The NMR spectrum (CDC13) showed absorption at 1.38 (s, 311), 1.60 (s, 3H), 1.8 (m, 21-1), 2.8 (m, 3H), 3.55 (dd, 1 H, J=4,91-1z), 4.1 (m, 1 H), 4. 43 (s, 1 H), 5.20 (s, 2H), 5.35 (61, 1 H, J=4Hz), 7.25 (s, 5H) and 7.4 (s, 5H) ppm.

C. benzyl 6P-[1(R(hydroxy-3-phenylpropyllpenicillanate sufflone To a solution of 585 mg. of benzyl 6p-[1-(R)hydroxy-3phenylpropyllpenicillanate in 35 ml. of methylene 55 chloride cooled to 0-5'C. was added 700 mg. of m-chloroperbenzoic acid, and the reaction mixture allowed to stir overnight. The solvent was removed under reduced pressure and the residue treated with an equal volume of water and ethyl acetate. The pH was adjusted to 7.2 with a saturated sodium bicarbonate solution and the organic layer separated. An equal volume of water was added to the ethyl acetate layer and the pH again adjusted as above to 8.4. The ethyl acetate layer was separate, washed with a saturated sodium 60 bicarbonated solution (3 x 50 mi.) and a brine solution, and was then dried over magnesium sulfate.

Removal of the solvent gave 678 mg. of the product as an oil which crystallized on standing, m.p. 142-143'C.

The NIVIR spectrum (CDC13) showed absorption at 1.30 (s, 3H), 1.6 (s, 311), 1.8 (m, 2H), 2.8 (m, 211), 3.83 (dd, 1 H, J=5,91-1z), 4.50 (s, 1 H), 4.55 (cl, 1 H, J=5Hz), 4.75 (m, 1 H), 5.2 (ABq, 2H), 7.2 (s, 511) and 7.38 (s, 5H) ppm.

16 GB 2 128 986 A 16 W. benzyl 6P-[1-(S)hydroxy-3-phenylpropyllpenicillanate sulfone The procedure of Example 11 C was followed, starting with 300 mg. of benzy] 60-[1 (S)hydroxy-3 phenyl propyllpenicil lanate and 361 mg. of m-chloroperbenzoic acid in 35 mi. of methylene chloride to give 346 mg. of product as an oil. 1 The NMR spectrum (C13C13) showed absorption at 1.28 (s, 3H), 1.52 (s, 1 H), 1.8 (m, 2H), 2.80 (m, 2H), 3.9 (dd, 5 1 H, J=5,101-1z), 4.45 (s, 1 H), 4.62 (m, 1 H), 4.67 (d, 1 H, J=51-1z), 5. 22 (ABq, 2H), 7.22 (s, 5H), and 7.38 (s, 51-1) ppm.

D. 6P-[1(R)hydroxy-3-phenylpropyllpenicillanic acid sufflone Benzyl 6p-[1 (R)hydroxy-3-phenylpropyllpenicillanate suifone (678 mg.) was added to a suspension of 700 1() mg. of prehydrogenated 5% palladium-on-calcium carbonate in 20 m]. of water-methanol OA). The mixture was shaken in a hydrogen atmosphere at an initial pressure of 52 psi for one hour. Atthattime an additional 700 mg. of catalystwas added and the hydrogenation continued for one hour. The catalyst was filtered and washed with water-methanol, The washings and filtrate were combined and the methanol removed in vacuo. The aqueous residue was extracted with ethyl acetate, followed by the adjustment of the pH of the aqueous to 1.5 and extraction with fresh ethyl acetate. The organic phase was backwashed with a brine solution and dried over magnesium sulfate. Removal of the solvent gave 304 mg., m.p. (138-140'C. dec. A sample of the free acid (190 mg.) was dissolved in ethyl acetate and was subsequently treated with 99 mg. of sodium 2-ethylhexanoate. After stirring overnightthe sodium salt of the desired productwas filtered and dried, 165 mg. 20 The NMR spectrum (DIVISO-D6) of the free acid showed absorption at 1.45 is, 1 H), 1.53 (s, 1 H), 1.8 (m, 2H), 20 2.80 (m, 2H), 3.85 (dd, 1 H, J=5,9Hz), 4.35 (s, 1 H), 4.60 (d, 1 H, J=51-1z), 4.75 (m, 1 H) and 7.23 (s, 51-1) ppm.

W. 6p-fl(S)hydroxy-3-phenylpropyllpenicillanic acid sufflone The procedure of Example 11 D was repeated starting with 346 mg. of benzyl 6p-[1 (S)hydroxy-3- phenyl propyll penicil la nate sulfone and 350 mg. of 5% palladium-on- calcium carbonate in 20 mi. of water-methanol (1:1) to give 196 mg. of the desired product, m.p. 146-148'C. dec.

A sample of 126 mg. of 6p[l (S)hyd roxy-3-phenyl propyll pen icil lanic acid suifone was dissolved in a small amount of ethyl acetate, and was subsequently treated with 57 mg. of sodium 2-ethylhexanoate. Some diethyl ether was added and the resulting precipitate was filtered and dried to give 57 mg. of the sodium salt of the desired product.

The NIVIR spectrum (DMSO-D6) of the free acid showed absorption at 1.47 (s, 1 H), 1.60 (s, 1 H), 2.0 (m, 2H), 2.8 (m, 2H), 3.9 (dd, 1 H, J=5,1 0Hz), 4.40 (s, 1 H), 4.67 (m, 1 H), 4.70 (cl, 1 H) and 7.2 (s, 51-1) ppm.

EXAMPLE 12

6p-[1 (R)Hydroxy-1 -(2'-pyridyi(methyllpenicillanic acid sulfone and 6p-[1 (S)Hydroxy-1 -(2'-pyridyi)methyll pen icil fa nic acid suifone A. benzyl 6P-[1(R) and (S) hydroxy- 1-(2'-pmethyll-penicillanate.

To a cooled (-78'C.) solution of 9.0 g. of benzyl 6,6dibromopenicillinate in 200 mi. of toluene under an argon atmosphere was added 11.8 mi. of t-butyl lithium and the resulting green solution allowed to stirfor 30 min. 2-Pyridylcarboxaldehyde (1.9 mQ was added and the reaction mixture stirred in the cold for45 min.

Subsequently 1.2 mi. of acetic acid was added in 25 mi. of diethyl ether over a period of 20 min. The mixture was allowed to stir in the cold for 30 min. and was then allowed to warm to 1 O'C. The reaction mixture was chromatographed on a Florisil column using toluene-ethyl acetate (2:1) as the eluent. Fractions (300 mi. 45 each) 3 thru 5 were combined and evaporated to give 4.8 g. of an oil.

The oil was dissolved in 60 mi. of dry benzene to which was then added 3. 2 mi. of tri-n-butyltin hydride.

The resulting reaction mixture was then refluxed under a nitrogen atmosphere for 2.5 hours. An additional 2.0 mi. of hydride was added and the heating continued overnight. The benzene was removed in vacuo and the residue slurried in hexane several times. The remaining oil was chromatographed on 500 g. of silica gel using toluene-ethyl acetate (2: 1) as the eluent.

Fractions 104thru 131 were combined and the solvent removed under reduced pressure to give 480 mg. of benzy] 6p-[1 (R)hyd roxy-1 -(2'-pyridyl)m ethyl] pen icil lanate as an oil. - The NIVIR spectrum (CDC13) showed absorption at 1.45 (s, 3H), 1.73 (s, 3H), 3.87 (dd, 1 H, J=4, 10Hz), 4.53 (s, 1WA.65 (M, 1H), 5.20 (m, 1H), 5.23 (s, 2H), 5.48 (d, 1H,J=4Hz), 7Ms, 5H), 7.5 (m, 31-1) and 8.6 (m, 11-1) ppm. 55 Fractions 136 thru 190 were combined and the solvent removed in vacuo to give 950 mg. of benzyl 6p-[1 (S)-hydroxy-1 -(2'-pyridyl) methyl] penici 1 lanate as an oil.

The NIVIR spectrum (CIDC13) showed absorption at 1.40 is, 3H), 1.68 is, 3H), 4.0 (m, 1 H, 4.05 (dd, 1 H, J=4.91-1z), 4.55 (s, 1 H), 5.2 (s, 2H), 5.22 (m, 1 H), 5.46 (cl, 1 H, J=4Hz), 7.3 is, 5H), 7,4 (m, 31-1) and 8.5 (m, 1 H) PPm.

B. benzyl 613-[1(R)hydroxy- 1-(2'-pyridyl)methyljpenicillanate suffione Under a nitrogen atmosphere 500 mg. of m-chloroperbenzoic acid was added to a solution of 480 mg. of benzyl 6p-[l (R)hydroxyl-1 -(2'pyridyl)methyllpenicillanate in 40 mi. of methylene chloride cooled to 05'C.

After stirring for one hour, the solent was removed in vacuo and the residue treated with an equal volume of 65 17 GB 2 128 986 A 17 water and ethyl acetate. The pH was adjusted to 7.2 with a saturated sodium bicarbonate solution followed by addition of sufficient sodium bisulfite until a negative starch-iodide test was given. After the aqueous layer was separated and fresh ethyl acetate was added, the pH was raised, as above, to 8.2. The ethyl acetate layer was separated, backwashed with a sodium bicarbonate solution and a brine solution and dried over magnesium sulfate. Removal of the solvent gave 480 mg. of an oil, which was subsequently chromatographed on 50 g. of silica gel using ethyl acetate as the eluent. Fractions 22 thru 55 were combined and the solvent removed in vacuo to give 125 mg. of the product.

The NIVIR spectrum (C1DC13) showed absorption at 1.22 (s, 3H), 1.50 (s, 3H), 4.40 (s, 1 H), 4.79 (m, 1 H), 4.80 (cl, 1 H, J=41-1z), 5.18 (ABq, 2H, J=121-1z), 5.6 (m, 1 H), 7.2 (m, 3H), 7.25 (s, 51-1) and 8.1 (m, 1 H) ppm.

B'benzyl 6p-[l(S)-hydroxy-l-(2'-pyridyl)methyllpenicillanate suffone Starting with 250 mg. of benzyl 6p-[l (S) hyd roxy-1 -(2'-pyridyl) methyl]pen ici Ilan ate and 320 mg. of m-chloroperbenzoic acid in 25 mi. of methylene chloride, and following the procedure of Example 12B, gave 240 mg. of the desired product as a white solid, m.p. 140-145T.

The NMR spectrum (CDC13) showed absorption at 1.23 (s, 3H), 1.59 (s, 3H), 4.6 (s, 1H), 4.8 (m, 2H), 5.3 (ABq,15 2H,J=12Hz), 5.95 (m, 11-10.4 (s, 51-10.5 (m,3H) and 8.4(m, 1H) ppm.

C. 6P-[1(R)hydroxy- 1-(2'-pyridyl)methyllpenicillanic acid sufflone To a suspension of 120 mg. of prereduced 5% palladium-on-calcium carbonate in 20 mi. of methanol-water W1) was added 120 mg. of benzyl 6P-[1(R)hydroxy-l-(2'- pyridyi)methyllpencillanate suifone, and the mixture was shaken in a hydrogen atmosphere at an initial pressure 47 psi for 30 min. An additional 120 mg. of catalyst was added and the hydrogenation continued for 45 min. at 47 psi. The catalyst was filtered, washed with methanol- water and the washings and filtrate combined. The methanol was removed in vacuo and the aqueous residue extracted with ethyl acetate. The aqueous layer was freeze dried to give 90 mg. of the desired product as the calcium salt.

The NIVIR spectrum (D20) of the calcium salt showed absorption at 1.50 (s, 3H), 1.65 (s, 3H), 4.35 (s, 1 H), 4.70 (m, 1 H), 5.18 (cl, 1 H, J=4Hz), 5.65 (cl, 1 H, J=1 1 Hz), 7.7 (m, 31-1) and 8.6 (broad cl, 1 H, J=Hz) ppm.

C'. 6P-[1(S)hydroxy- 1-(2'-pyridyl)methyllpenicillanic acid suffone The procedure of Example 12C was repeated, starting with 240 mg. of benzyl60-[l (S)hydroxy-l-(2'- 30 pyridyl)-methyll pencil lanate and 480 mg. of palladium-on-calcium carbonate in 20 ml. of methanol water, to give 170 mg. of the calcium salt of the desired product.

EXAMPLE 13 35 Starting with benzyi 6,6-dibromopenicillinate and the appropriate pyridylearboxaldehyde, and employing 35 the procedures of Example 12, the following compounds are prepared: 6p-[1 (R)hydroxy-1 -(3'pyridyl) methyl] penici 1 la nic acid suifone; 6p-[1 (R)hydroxy-1 (4'pyridyl) methyl] penici 1 lanic acid suifone; 6p-[1 (S)hyd roxylA -(4'pyridyl)-methyll pen icil lanic acid suifone; and 6p-[1 (S)hydroxy-1 -(3'pyridyl)methyllpenicillanic acid suffone.

EXAMPLE 14

6P-Acetoxymethylpenicillanic acid sulfone A. benzyl 6p-acetoxymethylpenicillanate sulfone To a solution of 500 mg. of benzyl 6p-hyd roxym ethyl penici I la nate sulfone a nd.196 m 1. of triethylam ine in 20 m 1. of methyl ene ch loride cool ed to 0-5oC. was added 0. 1 m 1. of acetyl ch loride a nd 10 mg. of 4-dimethylaminopyridine. After stirring for 20 min. the solvent was removed in vacuo and ethyl acetate was added to the residue. The resulting solids were filtered, and the filtrate washed successively with water, water at p 1.0, a saturated sodium bicarbonate solution and a brine solution. The organic phase was dried over magnesium sulfate and the solvent removed in vacuo to give 600 mg. of product as an oil.

Th NIVIR spectrum (CDC13) showed absorption at 1.30 (s,3H), 1.59 (s, 3H), 2-.1 (s,3H),4.2 (m, 1H),4.6 (s, 1H), 50 4.6 (m,21-10.65 (d, 1H,J=Hz), 5.22 (ABq, 2H,J=12Hz) and 7.4 (s, 5H) ppm.

B. 6p-acetoxymethylpenicillanic acid sufflone To a suspension of 600 mg. of 5% palladium-on-calcium carbonate, prereduced in 20 mi. of water-hydrogen to 50 psi for 20 min. was added 600 mg. of benzyi 6pacetoxymethylpenicillanic acid 55 suifone. The resulting mixture was shaken in a hydrogen atmosphere at an initial pressure of 50 psi for 45 min. The catalyst was filtered and washed with methanol-water. The filtrate and washings were combined and freeze dried to give 360 mg. of the desired product as a calcium salt.

The NMR spectrum (D20) of the calcium salt showed absorp tion at 1.5 (s, 3H), 1.61 (s, 3H), 2.18 (s, 3H), 4.25 (s, 1H),4.3 (m, 1H),4.60 (m, 21-1) and 5.07 (cl, 1H,J=4Hz) ppm.

EXAMPLE 15

60-Stearoyloxymethylpenicillanic acid sulfone A. benzyl 6p-stearoyloxymethylpenicillanate suffone Sta rting with 500 mg. of benzyl 6p-hyd roxym ethyl penici I lanate su Ifone, 430 mg. of stea royl ch loride,.19665 18 GB 2 128 986 A 18 mi. of triethylamine and 10 mg. of 4-dimethylaminopyridine, and following the procedure of Example 14A, there were obtained 784 mg. of the desired product as an oil.

The NMR spectrum (CIDC13) showed absorption at 1.4 (m 3H), 2.4 (m, 2H), 4. 2 (m, 1 H), 4.52 (s, 1 H), 4.60 (m, 2H), 4.63 (d, 1 H, J=41-1z), 5.22 (ABq, 2H, J=1 2Hz) and 7.4 (s, 5H) ppm.

B. 6p-stearoyloxymethylpenicillanic acid sulfone Following the procedure of Example 14B and starting with 776 mg. of benzy] 6pstearoyloxymethylpenicillanate sulfone and 880 mg. of 5% palladium-on-calcium carbonate in 25 m[. of methanol-water (1: 1) there was obtained 524 mg. of the calcium salt of the desired product. The calcium salt was subsequently suspended in 200 m]. of ethyl acetate and 200 m]. of water and treated with sufficient 6N 10 hydrochloric acid to give a pH of 2.0. The ethyl acetate layer was separated, backwashed with a brine solution and dried over magnesium sulfate. Removal of the solvent gave 260 mg. of the desired product as a white solid.

The NMR spectrum (CDC13 and DIVISO - D6) showed absorption at 1.4 (m, 3H), 2.35 (m, 2H), 4.2 (m, 1 H), 4.39 (s, 1H),4.60 (m, 21-1) and 4.63 (d, 11-1,J=4Hz) ppm.

EXAMPLE 16

Starting with the appropriate acid chloride and requisite benzyl 6phydroxyalkylpenicillanate sulfone and employing the procedure in Example 14A and B, the following compounds are prepared:

6p-[1 (R)acetoxy-n-butyllpenicillanic acid sulfone; 6p-[1 (S)acetoxy-3-m ethyl-n-butyl]pen ici 1]an ic acid sul- 20 fone; 6p-[1 (S)acetoxy-2-phenethyllpenicillanic acid suffone; 6p-[1 (S) acetoxy- 1 -(3'-pyridyl) methyl pen ici 11 a nic acid su ifo ne; 60-n-butyryloxym ethyl pen ici 11 an ic acid sulfone; 6p- [1 (S)-s-butyryloxy-n-penicillanic acid sul fone; 6P-[(S)-n-butyryloxy-2-m ethyl-n-p ro pyl]-pen ici 11 an ic acid sulfone; 6p-[1 (S)-n-butyryloxybenzyil penicillanic acid sulfone; 6p-[1 (R)-n-butyryl oxy-1 -(2'-pyridyf) m ethyl] penici 11 an ic acid suffone; 6p-[1 (S)-s butyryloxy-1 -(4'-pyridyl)methyll penici 1 fan ic acid su Ifone; 6p-nvaleryfoxymethyl penicillanic acid sulfone; 25 6P-[1(S)-n-valeryloxyethyllpenicillanic acid suffone; 6p-[1(R)-ivaleryloxy-n-pentyllpenicillanic acid sulfone; 6p-[1 (R) trimethylacetoxy-2,2-dimethyl-n-propyilpenicillanic acid sulfone; 6p-[1 (R)-n-valeryloxy-3 phenyl propyll penici 11 an ic acid sulfone; 6p-[1 (R)-i-va 1 eryl oxy-1 - (2'-pyridyl) methyl] pen ici 1 la nic acid sulfone; 6p-[1 (S)-tri methyl acetoxy-1 -(4'-pyridyl) methyl] penici 1 lanic acid suifone;6p-octanoyloxymethylpenicillanic acid sulfone; 6p-[1 (R)octanoyloxy-n-pentyllpenicillanic acid sulfone; 6p- [1 (S)octanoyloxy-2 phenethyllpenicillanic acid sulfone; 6p-[1 (S)octanoyfoxy-1 -(3'-pyridyl) methyl] pe nici 1 fan ic acid sulfone; 6p-[1 (S)hendecanoyloxy-2-methyl-n-propyllpenicillanic acid sulfone; 6p- [1 (R)hendecanoyloxybenzyi] peniciflanic acid sulfone; 6p-[1 (S)hendecanoyloxy-1 -(4'- pyridyi)methyllpenicillanic acid sulfone; 6p paimitoyloxymethylpenicillanic acid sulfone; 6p-[1 (S)paimitoyloxyethyllpenicillanic acid sulfone; 6p [1(R)paimitoyloxy-2-phenethyllpenicillanic acid sulfone; 6P[1(S)paimitoyloxy-l-(3' pyridyl)methyll pen icil 1 an ic acid su Ifone; 6p-[1 (S)pal mitoyloxy-1 - 4'-pyridyl)methyll penicillanic acid sulfone; 6p-[1 (R)stea royloxyethyll penicil fan ic acid su Ifone; 6p-[1 (S)stearoyfoxy-2,2-dimethyi-n-propyllpenicillanic acid sulfone; 6P-[1(R)stearoyloxy-3-phenylpropyll-penicillanic acid sulfone; 6P-[1(S)stearoyloxy-l-(2' pyridyi)methyll pen icil fan ic acid su Ifone; and 6p-[1 (S)stearoylA - (4'-pyridyi)methyll penicil fan ic acid sulfone.

EXAMPLE 17

6p-Benzoyioxymethylpenicillanic acid sulfone A. benzyl benzoyloxymethylpenicillanic acid sulfone To a solution of 300 mg. of benzyl 6p-hydroxymethylpenicillanate sulfone and 0.11 mi. of triethylamine in 25 ml. of methylene chloride cooled to O-WC. was added 0.094 mi. of benzoyl chloride and 10 mg. of 4-dimethylaminopyridine. After stirring in the cold for 30 min. the solution was washed successively with water, water at pH 1.0, a saturated sodium bicarbonate solution and a brine solution. The organic phase was dried over magnesium sulfate and the solvent removed in vacuo. The residue was chromatrographed on 20 g. of silica gel using toluene-ethyl acetate (8:1) as the eluent. Fractions 15 thru 30 were combined and concentrated in vacuo to give 280 mg. of the product as an oil.

The NMR spectrum (CDC13) showed absorption at 1.26 (s, 3H), 1.53 (s, 3H), 4.2 (m, 1 H), 4.57 (s, 1 H), 4.79 (d, 1 H, J=4Hz), 4.9 (m, 2H), 5.2 (ABq, 2H, J=1 2Hz), 7.4 (s, 5H), 7.5 (m, 31-1) and 8.2 (m, 21-1) ppm B. 6p-benzoyloxymethylpenicillanic acid sulfone To a suspension of 270 mg. of prereduced 5% pal ladium-on-calciu m carbonate in 15 mi. of water-methanol 55 (1:1) was added 270 mg. of benzyl 6p-benzoyloxymethylpenicillanic acid sulfone, and the resulting mixture was shaken in a hydrogen atmosphere at an initial pressure of 50 psi for 40 min. The catalyst was filtered and the methanol was evaporated. The residual aqueous was extracted with ethyl acetate and was then freeze dried to give the calcium salt of the product, 200 mg.

The NMR spectrum (D20) of the calcium salts showed absorption at 1.5 (s, 3H), 1.6 (s, 3H), 4.8 (m, 3H), 5.1 60 (d, 1 H, J=41-1z), 7.6 (m, 31-1) and 8.0 (m, 21-1) ppm.

19 1 GB 2 128 986 A 19 EXAMPLE 18

Starting with the appropriate benzyl 6p-hydroxyalkylpenicillanate sulfone and requisite benzoyl chloride, and employing the procedures of Example 17A and B, the following esters are prepared:

?- H - 5 c 0 -. 0 2,, CH3 1 X \ _ - ' S.. CH 3 R 4 0 'CO 2H 10 X R4 4-ClH- 15 2-F- H - 4-CH3- H 4-Cl- CH3- (S) 4-Cl- CH3- (R) 20 4-CF3- CH3- (R) 4-F- C1-13(C1-12)2_ (R) 2-CH3- CH3WI-12)2_ (R) 3-CH30- CH3WH2)2_ (R) 2-FCH3CH(CH3WI-12- (S) 25 H- C1-13(C1-12)2_ (R) H- (CH3)3C- (S) 3-Cl(CH.3)3C_ (R) H- C61-15- (S) 3-CH3- C6H5- (S) 30 4-CH30- C61-15- (R) 2-FC6H5CH2- (S) 2-CH3- C61-15(CHA2- (R) 4-F- C6H6WHA2- (R) 4-CF3C61-15(CHA2- (S) 35 4-CH3- C61-15(CH2)2_ (S) H- 2'-C51-14N- (R) 3-CH32'-C51-14N- (S) 4-CF3- 2'-C51-14N- (R) H- 3'-Crffi4N- (S) 40 4-CH30 X-CMN(S) 4-CH3 X-C51-14N- (S) 3-F- X-CMN- (R) H- 4'-C5H4N- (R) 2-F- 4'_C51-14N(R) 45 4-CF3- 4'-C51-i4N- (S) 4-CH3- 4'_C51-14N- (S) EXAMPLE 19 50 6p-4'-Aminobenzoyloxymethylpenicillanic acid sulfone A. benzyl 6p-4'-nitrobenzoyloxymethylpenicillanate sufflone Under an argon atmosphere 264 mg. of 4'-nitro-benzoyl chloride and 10 mg. of 4-dimethyiamirlopyridine was added to 500 mg. of benzyl 6phydroxymethylpenicillinate sulfone and 0.196 mi. of triethylamine in 20 mi. of methylene chloride cooled to O-WC,After stirring in the cold for 30 min. the reaction mixture was washed successively with water, water at pH 1.0, a saturated sodium bicarbonate solution and a brine solution. The organic phase was dried over magnesium sulfate and the solvent removed in vacuo to give 657 mg. of the product as a semi-solid.

The NIVIR spectrum (CDC13) showed absorption at 1.33 (s, 3H), 1.58 (s. 3H), 4.3 (m, 1 H), 4.58 (s, 1 H), 4.8 (cl, 1 H, J=41-1z), 4.9 (m, 2H), 5. 23 (ABq, 2H, J=1 2Hz), 7.39 (s, 51-1) and 8.2 (s, 41-1) ppm.

B. 6P-4'-aminobenzoyloxymethylpenkillanic acid sulfone To a.suspension of,650 mg. of preredLLced,5% palladium-on-calcium carbonate in 20 ml. of water-methanol (1:1) and 10 ml. of tetrahydrofuran was added 650 mg. of benzyl 60-4'-n itro benzoyl oxym ethyl pen ici Ilan ic acid sulfone, and the resulting mixturewas shaken in a hydrogen atmosphere atan initial pressure of 50 psi for one hour. The spent catalyst was filtered andrthe residue partitioned between ethyl acetate and water.

GB 2 128 986 A the aqueous layer was separated and freeze dried to give 560 mg. of the product as the calcium salt.

The NMR spectrum (D20) of the calcium salt showed absorption at 1.5 (s, 3H), 1.6 (s, 3H), 4.39 (s, 1 H), 4.70 (m, 3H), 5.1 (d, 1 H, J=41-1z), 6. 78 (d, 2H, J=9Hz) and 7.8 (d, 2H, J=9Hz) ppm.

EXAMPLE 20

The procedures of Example 19 are repeated, starting with the requisite benzyl 6phydroxyalkylpenicillanate suifone and appropriate nitrobenzoyl chloride, to give the following compounds as their calcium salts:

H 0 2, CH 0 - 2 S CH 3 N N 4 0 C02 H NH 2 NH2 Ring Position R4 3- H 2- H 2- CH3- (R) 25 4- CH3- (R) 4- CH3- (S) 2- C1-13(CH2)2_ (R) 3- C1-13(CHA2(R) 3- CH3CH(CH3)CH2- (S) 30 4- CH3CH(CH3)CH2- (S) 2- (CH3)3C- (R) 3(CH36C- (S) 3- C61-15- (R) 4- C61-15- (R) 35 2- C6H5CH2- (S) 3- C6HrCH2(S) 4- C61-15CH2- (S) 2- C61-15(C1-12)2_ (R) 4- C6H5(CHA2- (R) 40 22-C51-14N- (R) 4- 2-C5H4i\11- (R) 3- 3-C51-i4N- (S) 3- 4-C51-14N- (S) 44-C51-14N- (R) 45 EXAMPLE 21

6p-4'-Tolylsulfonyloxymethylpenicillanic acid sulfone A. benzyl 6P-4'-tolylsulfonyloxymethylpenicillanate To 1.24 g. of 4-tolysulfonylchloride in 3.5 mi. of pyridine cooled to OOC. and under an argon atmosphere, 50 800 mg. of benzyl 6p-hydroxymethylpenicillanate in 1.5 mi. of pyridine was added dropwise. After stirring in the cold for 2 hrs., 0.80 mi. of water was added and the stirring continued for 30 min. at OOC. The reaction mixture was added to 30 ml. of water and the pH adjusted to 1.0 with dilute hydrochloric acid. The aqueous was extracted with diethyl ether and the organic phase was separated and washed successively with 1.2N hydrochloric acid, water and a brine solution. The organic layer was dried over magnesium sulfate and evaporated to an oil, 841 mg., which was chromatographed on 100 g. of silica gel using chloroform-ethyl acetate (10:1) as the eluent.

Fractions 10 thru 25 were combined and the solvent removed in vacuo to give 680 mg. of the product.

B.6P-4'-tolylsulfonyloxymethylpenicillanic acid 60 To a suspension of 680 mg. of prereduced 5% palladium-on-calcium carbonate in 20 ml. of methanol-water (1:1) was added 680 mg. of benzyl 6P-4'tolyisulfonyloxymethylpenicillanate, and the reduction continued at 49 psi for 30 min. An additional 680 mg. of catalyst was added and the reaction continued for an additional 30 min. The catalyst was filtered and the methanol evaporated from the filtrate. The aqueous residue was extracted with ethyl acetate and the aqueous layer acidified to pH 2.0. Fresh ethyl acetate was used to extract 65 21 GB 2 128 986 A the acidified aqueous. The organic phase was separated dried over magnesium sulfate and the solvent removed in vacuo to give 463 mg. of the product as a semi-solid.

* The NMR spectrum (CDC13) showed absorption at 1.57 (s, 3H), 1.6 Is, 3H), 2.37 (s, 3H), 4.1 (m, 3H), 4.2 (s, 1 H), 5.4 (d, 1 H, J=4Hz), and 7.6 (ABq, 4H, J=1 OHz) ppm.

C. 6p-4'-tolylsulfonyloxymethylpenicillanic acid sufflone Water (20 mi.) was added to a solution of 460 mg. of 6p-4'-to lyisu Ifonyl oxym ethyl pen icil 1 an ic acid in 20 mi. of methylene chloride and the pH of the resulting mixture adjusted with a sodium hydroxide solution to 6.9. The aqueous layer was separated and the organic layerfurther extracted with water (2 X 20 mi.). To the combined water layer and washings was added dropwise 238 mg. of potassium permanganate in 5 mi. of 10 water containing 0.16 mi. of phosphoric acid. During the reaction period (45 min.) the pH of the reaction was maintained at 6.0-6.4 by the addition of aqueous sodium hydroxide. The pH of the reaction mixture was then adjusted to 1.5 with 6N hydrochloric acid and 20 mi. of ethyl acetate was added. After cooling the mixture to O'C., 460 mg. of sodium bisulfite was added in one portion. The pH was readjusted to 1.5 with 6N hydrochloric acid and the organic phase was separated, backwashed with a brine solution and dried over 15 magnesium sulfate. Removal of the solvent gave 300 mg. of the product as a foam.

The NMR spectrum (CDC13) showed absorption at 1.45 (s, 3H), 1.65 (s, 311), 2.45 (s, 31-1), 4.4 (m, 31-1), 4.42 (s, 1 H) 4.8 (d, 1H, J=41-1z) and 7.6 (ABq, 4H, J=MHz) ppm.

22 GB 2 128 986 A 22 EXAMPLE 22

Starting with the appropriate sulfonyl chloride and requisite benzyi 6phydroxyalkylpenicillanate, and employing the procedures of Example 21, the following compounds are prepared:

H H 0 5 2 CH 3 R -CH 3 0 1 CH3 R4 0 C02 H 10 R3 R4 CH3S02- H (C1-13)2CHS02- H- 15 C1-13(C1-12)3S02- H CH3S02- CH3- (S) CH3S02(CH3)2CHCH2- (S) CH3CH2S02- C6H5- (R) CH3S02- 2'-C51-114N- (S) 20 C1-13(C1-12)2S02- 4'-C51-14N- (S) C61-15S02- C61-15(C1-12)2_ (S) C61-15S02- CH3WI-12)2_ (R) C61-15S02- H 2-C3C61-14S02- H4-CH3C61-14S02- CH3- (R) 2-CH3C61-14S02- C1-113(CH2)3_ (R) 3-CH3C61-14S02C61-1r)(CHA2- (S) 4-CH3C61-14S02- X-C51-14N- (S) 4-CH30C61-14S02- H4-CH30C61-14S02- (CH3)2CH- (S) 3-CH30C61-14S02- C61-15- (S) 2-CH30C61-14S02- C21-15- (R) 4-CH30C61-14S02- 4'-C51-14N- (S) 2-17C61-14S02- H- 35 4-FC61-14S02- CH3- (S) 3-FC61-14S02- (CH3)3C- (R) 4-Kr,H4S02- C61-15CH2- (S) 2-FCr)H4S02- 2'-Cr)1-14N- (S) 2-CIC61-14S02- H40 2-CIC61-14S02- (CH3)2CHCH2- (S) 4-CIC61-114S02- C61-16CH2- (R) 4-CIC61-14S02- Crl-15(CHA2- (R) 4-CIC61-14S02- 4'-Cr)1-14N- (S) 2-BrC61-14S02- H- 45 4-BrC61-14S02- CH3- (R) 4-BrC61-14S02- C6H5- (S) 3-CF3C61-14S02- H 3-CF3C61-114S02- C21-15- (R) 4-CIF3C61-14S02- H4-CF3C61-14S02- CH3- (S) 4-CF3C61-114S02- C61-15CH2- (S) 4-CIF3C61-14S024'-C51-14N- (R) EXAMPLE 23

6P-Sulfooxym ethyl penici I I an ic acid sulfone A. benzyl 6p-sulfboxymethylpenicillanate sulfone pyridinium saft To a solution of 953 mg. of benzyl 6p-hydroxymethylpenicillanate sulfone in 75 ml. of dimethylacetamide under a nitrogen atmosphere is added 860 mg. of sulfur trioxide-pyridine complex and the reaction mixture allowed to stir for 45 min. at room temperature, The mixture is stirred into 400 mi. of hexane and allowed to 60 stir for 30 min. The hexane is decanted and fresh hexane added. This procedure is repeated twice to give 900 mg. of the desired product.

B. 6p-sulfboxymethylpenicillank acid suffione A mixture of 532 mg. of benzyl 6p-su Ifooxym ethyl penicil Ian ate sulfone pyridiniurn salt in 10 mi. of water 65 23 GB 2 128 986 A 23 containing 174 mg. of sodium bicarbonate is added to a suspension of 500 mg. of prereduced 10% pal ladiu m-on-charcoa I in 25 ml. of water, and the resulting mixture is shaken in a hydrogen atmosphere at 50 psi for one hour. An additional 500 mg. of catalyst is added and the reaction continued for 30 min. A final 500 mg. of catalyst is made and the reduction continued for 30 min. The catalyst is filtered and washed with water. The filtrate and washing are combined and freeze dried to give the crude product, which is purified by chromatographing on sephedex using water as the eluent. The fractions containing the product are combined and freeze dried to give the purified product.

EXAMPLE 24

Starting with the appropriate benzyl 6p-hydroxyal kyl pen icil lanate suifone and employing the procedures 10 of Example 23A and B, the following compounds, as their disodium salts are prepared.

H H u2 CH HO 0 S 3 S -CH 4() R C.3 C02H R4 20 CH3- (S) CH3-(R) CH3CH2- (S) CH3WHA2- (R) 25 C1-13(CH2)3_ (S) C1-13(CHA3- (R) CH3CH(CH3)- (S) (CH3)3C(R) (CHINC- (S) 30 C61-15- (S) C61-15- (R) C61-15CH2- (S) C61-15(CHA2- (S) C61-15(CH2)2_ (R) 35 2'-C51-14N- (S) 4'_C51-14N- (S) 40 EXAMPLE 25 Piva loyl oxym ethyl 6p-hydroxymethylpenicillanate sulfone To a solution of 1.0 g. of 6p-hydroxymethylpenicillanic acid sulfone sodium salt in 10 mi. of dimethylformamide and cooled to 0-5'C. was added to 0.52 mi. of chloromethyl pivalate. After stirring overnight at room temperature, the reaction mixture was poured into a mixture of water- ethyl acetate. The ethyl acetate layerwas separated, backwashed with water (3 x 100 ml.) and a brine solution (3 X 50 ml.) and dried over magnesium sulfate. The solvent was removed in vacuo to give 1.1 g. of the product as an oil.

The NIVIR spectrum (CDC13) showed absorption at 1.27 (s, 9H), 1.42 (s, 3H), 1.6 (s, 3H), 2.9 (bs, 1 H), 4.2 (m, 3H), 4.58 (s, 1 H), 4.75 (m, 1 H) and 5.82 (ABq, 2H, 8A_8Bl 6Hz) ppm.

24 GB 2 128 986 A 24 EXAMPLE 26

Employing the procedure of Example 25r and starting with the appropriate 6p-hydroxyalkylpenicillanic acid sulfone or derivative thereof and the requisite halide, the following esters are prepared: H H 5 2 CH 3 S R CH3 30-CH A -:_r C02R, 10 R3 R4 R, H- H- - -CH(C1-13)02CCH3 H- H- - -CH202CCH3 15 H- H- - -CH(CH3)02C(CH2)4CH3 H- CH3- (S) -CH202CCH3 H- CH3- (R) -CH202COCH(C1-13)2 H- (CH3)2CH - (S) -CH202CC(CH3)3 H- C61-16- (R) -CH202CC1-13 20 H- C61-15- (R) -C51-1302a H- C61-15CH2- (R) -CH202COCH3 HC61-15CH2- (S) -C4H502 b H- C61-IB(C1-12)2_ (S) -CH(CH3)02COC2H5 HC61-15(C1-12)2_ (R) -C81-1502C 25 H- T-C5H4N- (R) -CH202CCH3 H- T-C51-14N(S) -CH(C1-13)02COC2H5 H- X-C51-14N- (S) -C(C1-13)202CO(CH2)2CH3 H3'-C5H4N- (R) -C41-1302 a H- X-C51-14N- (R) -CH202COCH3 30 H- 4'-C51-14N(S) -CH(CH3)02C(CH2)4CH3 H- 4'-Cr)H4N- (S) -(CH3)202CC(CH3)3 H- 4'-C^N(R) -CH(CH3)02CCH3 H- 4'-C51-14N- (R) -C,H,02c CH3CO- CH3(C1-12)2- (R) -CH202COCH(CH3)2 35 CH3CO- (CH3)2CH2- (S) -CH202CCH3 CH3CO- C61-16(CH2)2(S) -C4H502 b CH3CO- X-C51-14N- (S) -CH(CH3)02CO(CH2)3CH3 (C1-13)2CHCOC2H5- (S) -CH202C(CH2)4CH3 C1-13(C1-12)2CO- (CH3)2CH- (S) -CH(C1-13)02CCH3 40 C1-13(C1-12)2CO- C61-15- (S) -CH(CH3)02CCH3 C1-13(C1-12)2CO- T-CMN(R) -CH202CC1-13 C1-13(C1-12)2CO- 4'-C51-14N- (S) -CH202CCH(CH3)2 CH3WH2)3CO- H- - -CH202CC1-13 CH3WH26CO- H- - -CH(C1-13)02CO(CH2)2CH3 45 (CH3)2CHCH2CO- CH3WII2)3- (R) -CH202CC(CH3)3 (C1-13)2CHCH2CO- T-C51-14N(R) -C4H302 a (CH36CCO- 4'-C51-14N- (S) -CH(CH3)02CCH3 CH3WH2)6CO- H- -C(CH3)202MCH3)3 C1-13(C1-12)6CO- C6HsCH2- (S) -CH202CC1-13 50 CH3WH2)6CO3'-C51-14N- (S) -C.1---1502c C1-13(C1-12)9CO- H- - -CH(CH3)02CCH3 CH3(CH2)9CO- H- - -CH(C1-13)02COC2H5 CH3(CH2)9CO- (CH3)2CH (S) -CH202COCH(C1-13)2 C1-13(C1-12)14CO- H- - -CH202CCH3 55 C1-13(C1-12)14COH- - -CH202C(CH2)4CH3 C1-13(C1-12)14CO- CH3- (S) -CH(CH3)02COCH3 C1-13(C1-12)14CO- CH3- (S) -C41-1502 b C1-13(C1-12)14CO- 3'-C51-14N- (S) -CH202COCH3 C1-13(C1-12)16CO- H- - -CH202CC(CH3)3 60 CH3WH2)16CO- H- -CH(CH3)02COC2Hrs a 4-crotonolactonyl by-butyrolacton-4-yl c3-phthalidyl 65 iz z GB 2 128 986 A 25 EXAMPLE 27 Acetoxymethyl 6p-[l (S)methylsulfonyloxy-2-phenethyll- penicillanate sulfone To a solution of 2.22 g. of 6p-[l (R)ethylsufl onyl oxybenzyll pen ici I [an ic acid sulfone sodium salt in 30 ml. of dimethy1formamicle and cooled to 5'C. is added 648 mg. of chloromethyl acetate. The reaction mixture is allowed to stir at room temperature overnight, and is then poured into water-ethyl acetate. The ethyl acetate layer is separated, backwashed with water and dried over magnesium sulfate. Removal of the solvent in vacuo gives the desired product.

EXAMPLE 28

The procedure of Example 27 is employed, starting with the appropriate 6psulfonyloxyalkylpenicillanic 10 acid sulfone and halide, to give the following products:

H H 1 0 CH 3 R S H 30-Ce H C 3 R 7 n 4 0 CO 2R1 R3 R4 R, CH3S02- H- -CH202CC113 CH3S02- CH3 (S) -CH(CH3)02COC2115 25 CH3CH2S02 C611s- (R) -CH202COCH3 (CH3)2CHS02- H- - -C(CH3)202CO(CH2)2CH3 C113(CH2)2S024'_C5H4N- (S) -CH202CC(CH3)3 CF13(C1126S02- H- - -C(CH3)202CC(CH3)3 2-CH3C6H4S02- H- - _C4H302 a 30 4-CH3CE;H4S02- CH3- (R) -CH202CCH(CH3)2 2-CH30C6H4S02- C2F15- (R) -CH(CH3)02COCH3 3-CH30C6H4S02- C6H5- (S) -CH202CCH3 4-CH30C6H4S02- 4'-C5H4N- (S) -CH202CCH3 2-FC61-14S02- H- -CH202CCH3 35 4-FC6114S02- CH3- (S) -CH202CC(CH3)3 2-CIC6H4S02(CH3)2CHCH2- (S) - C(CH3)202C0 (CHA2CH3 4-CIC6114S02- C6H5CH2- (R) -CH202CCH3 4-CIC61H4S02- C6115WHA2- (R) -CH202COCH3 2-BrC6H4S02- H- _C4H502 b 40 4-BrC6H4S02- CH3- (R) -C8H502C 3-CF3C6H4S02- C2115- (R) -C(C1-13)202CCH3 4-CF3C6H4S02 CH3- (S) -CH202CO(CH2)2CH3 4-CF3C6H4S02C6H5C112- (S) -CH202COCH(CHA2 C6H5S02- C1-13(CHA2- (R) -CH2COCH(CHA2 45 C611502- H- - -CH202CC(CH3)3 C6H5S02- C61H5(CHA2- (S) -CH(C113)02CCH3 NaSO3- H- - -CH202CCH3 NaS03- CH3(CHA2- (R) -CH(CHA02COC2H5 NaS03Ce)H5C1-12- (S) -CH202CC(CH3)3 50 NaS03- 2'-CsH4N- (5) -CH202COC2Hr) 4-crotonolactonyl ^-butyrolacton-4-y.1 '3-phthalidyl 55 EXAMPLE 29

Piva loyl oxym ethyl 6P-Benzoyl oxym ethyl pen ici I I a nate sulfone To a solution of 1.95 g. of 6p-benzoyloxymethylpenicillanate sulfone sodium salt in 25 ml. of dry dimethylformamide cooled to 0-5'C. is added 900 mg. of chloromethyl pivalate and the mixture allowed to 60 stir overnight at room temperature. The mixture is then poured into a mixture of ethyl acetate-water. The ethyl acetate layer is separated, backwashed with water and a brine solution and dried over magnesium sulfate. Removal of the solvent in vacuo gave the desired product.

26 GB 2 128 986 A EXAMPLE 30

Starting with the appropriate 6p-hydroxyalkylpenicillanic acid sulfone derivative as the sodium salt and the requisite alkyl halide, and employing the procedure of Example 29, the following compounds are prepared:

0 0 CH 3 -C-O-C S C H AHA ono" I- N' 3 X 0 co 2 R 1 X R4 R, 4-Cl- H- -CH202CCH3 4-Cl- CH3- (R) -CH(CH3)02CCH3 3-Cl- (CH3)3C- (R) -CH202MCH3)3 2-F- H- - -CH(CH3)02COC2H5 4-F- CH3WH2)2_ (R) -CH202MCH3)3 3-17- X-C5H4N(R) -C(C1-13)202CCH3 3-Br- (CH:3)2CHCH2- (S) -C41-11302a 4-Br- C61HI5CH2(R) -CH202COCH3 2-CH3- CH3WI-12)2_ (R) -CH(CH3)02COC21H5 3-CH3- C6H5- (S) -CH202C(CH2)4CH3 4-CH3- H- - -C81-1502C 4-CH3- C61-115(CH2)2_ (S) -CH202(CH24CH3 4-CH3- 4'-C5H4N- (S) -C(CH3)202COCH3 H- 4'-C5H4N- (S) -CH202CCH3 H- CH3(CH2)2_ (R) -CH(CH3)02CCH3 4-CIF3- CH3- (R) -C41-1502 b 4-CF3 C^(C1-112)2_ (S) -CH202COC21H15 3-CH30- CH3WH2)2_ (R) -CH202MCH3)3 4-CH30- C6H5- (R) -CH(CH3)02COC2H5 4-CH30- 3'-C5H4N- (S) -C(CH3)202C(CH2)2CH3 4-crotonolactonyl 6-y-butyrolacton-4-yi '3-phthaiidyl EXAMPLE 31 Pivaloyloxymethyl 6p-4'-a m i no benzoyloxymethyl pen ici 11 a nate sulfone A. pivaloyloxymethyl 60-4'nitrobenzoyloxymethylpenicillanate sufflone To a cold solution of (0-50C.) of 1.89 g. of piva Icy] oxym ethyl 6p- hydroxymethylpenicillanate sulfone---695 mi. of triethylamine and 10 mg. of 4-dimethylaminopyridine in 60 mi. of methylene chloride under an atmosphere of argon is added 1.11 g. of 4'-nitrobenzoyl chloride. The resulting reaction mixture is allowed to 45 stir at room temperature for 2 hrs. and is then extracted with cold water. The organic layer is separated and washed successively with water at pH 1.0, a saturated sodium bicarbonate solution and a brine solution. The organic layer is then dried over magnesium sulfate and concentrated to dryness in vacuo to give the desired intermediate.

26 B. pivaloyloxymethyl 6P-4'-aminobenzoyloxymethylpenicillanate suffione To a solution of 500 mg. of piva I oyloxym ethyl 6P-4'-n itro benzoyl oxymethyl pen ici I I anate sulfone in 35 mi. of climethoxyethane is added 250 mg. of 5% palladium-on-charcoal and the resulting suspension shaken in a hydrogen atmosphere at an initial pressure of 30 psi. After 3 hrs. the spent catalyst is filtered and the filtrate concentrated in vacuo to give the desired product.

3 1 27 GB 2 128 986 A 27 EXAMPLE 32 Starting with the appropriate nitrobenzoyl chloride and 6p- hydroxyalkyl pen icillan ic acid ester suifone and employing the procedure of Example 31A and B, the following compounds are prepared:

0 2 CH 3 S CH c 0 Cli-ii 3 R R NH2 4 0 C021 NH2 Ring Position R4 R, 2 H- -CH202CCH3 15 4 H- - -CH202COCH3 2 CH3- (S) -CH202CCH3 3 (CH3)2CH- (S) -CH202CC(CH3)3 4 C6H5- (R) -C5H302 a 4 C6H5C1-12- (S) _C41H1502 b 20 4 CM(CH2)2_ (S) -CH(CH3)02COC2Hs 3 C6115(CHA2- (R) -CBH592C 2 2'_C5H4N- (R) -CH202CCH3 4 4'-C51-14N- (S) -CH(CH3)02C(CHA4CH3 3 3'_C51-14N (R) -CH202COCH3 25 a4-crotonolactonyl b -y-butyrolacton-4-yl '3-phthalidyl EXAMPLE 33

6P-Methoxycarbonyloxymethyl pen icil lanic acid sulfone A. benzyl 6p-methoxycarbonyloxymethylpenicillanate suffione To a solution of 500 mg. of benzyl 60-hydroxymethylpenicillanate sulfone and.1 96 ml. of triethylamine in 20 mi. of methylene chloride cooled to O'C. is added.1 ml. of methyl chloroformate and 10 mg. of 4-dimethylaminopyridine, After stirring for 20 min. the solvent is removed under reduced pressure and ethyl acetate is added to the residue. The resulting solids are filtered, and the filtrate washed successively with water, water at pH 1.0, a saturated sodium bicarbonate solution and a brine solution. The organic phase is dried over magnesium sulfate and the solvent removed in vacuo to give the desired intermediate.

B. 6p-methoxycarbonyloxymethylpenicillanic acid sulfone To a suspension of 600 mg. of 5% palladium-on-calcium carbonate, prereduced in 20 ml. of water with hydrogen at 50 psi for 20 min. is added 600 mg. of benzyl 6pmethoxycarbonyloxymethylpenicillanate sulfone. The resulting mixture is shaken in a hydrogen atmosphere at an initial pressure of 50 psi for 45 min.

The catalyst is filtered and washed with methanol-water. The filtrate is treated with 75 ml. of ethyl acetate 45 and the pH adjusted to 1.5 with 6N hydrochloric acid. The organic phase is separated, dried over magnesium sulfate and concentrated in vacuo to give the final product.

28 GB 2 128 986 A EXAMPLE 34

Starting with the requisite benzy] 6p-hydroxyalkylpenicillanate sulfone and alkyl chloroformate, and employing the procedures of Example 33A and B, the following compounds are prepared:

28 H H 0 CH 3 R 3 O-CH S CH 3 4 0 N co 2 H F33 R4 C21-150CO- CH3- (S) 15 CH30CO- CH- (R) CH30CO- C2H5- (R) C1-13(CH2)20CO- CH3(CH2)2_ (R) (CH3)2CHOCO- (CH3)2CH- (S) CH30CO- C1-13(CH2)4_ (S) 20 C1-13(C1-12)2_ (CH3)3C- (R) CH30CO- C61-1s- (S) C2H50CO- C6H5CH2- (S) (CH3)2CHOCOCrH5C1-12- (R) CH30CO- C61-15(CH2)2_ (S) 25 CH30CO- 2'-C51-14N- (R) CH30CO- 2'-C51-14N- (S) C21-150CO- E'-C51-14N- (R) CH(CH2)20CO- 3'-C^N(S) C21-150CO- 4'-C5i---14N- (S) 30 CH30CO- 4'_C5i---14N- (R) EXAMPLE 35

Acetoxymethyl 6p-m eth oxyca rbonyl oxymethyl pen ici I I anate su Ifone To a solution of 1.6 g. of 6p-methoxycarbonyloxymethyl pen icillanic acid sulfone in 20 ml. of dry dimethylformamide at 50C. and under a nitrogen atmosphere is added 120 mg. of sodium hydride. After stirring for 10 min. 648 mg. of chloromethyl acetate is added and the reaction mixture allowedto stir overnight at room temperature. The reaction mixture is poured into a mixture of water-ethyl acetate and the organic phase is subsequently separated, backwashed with water and a brine solution and dried over 40 magnesium sulfate. Removal of the solvent in vacuo gives the final product.

11 f 29 GB 2 128 986 A 29 EXAMPLE 36 The procedure of Example 35 is employed, starting with the appropriate 6palkoxycarbonyloxyalkylpenicillanic acid sulfone and alkyl chloride, to give the following compounds:

H H 0 2 CH 3 S R 3 0 CH [I - 7 CH 3 4 0 C02R1 R3 R4 R, 15 C21-IE;0CO- CH3(S) -CH202CCH3 CH30CO- C2H5- (R) -CH(CH3)02CCH3 CH3(C1-12)20CO- CH3(CH2)2_ (R) -CH202COC2H5 (CH3)2CHOCO- (CH3)2CH- (S) -CH(CH3)02CO(CH26CH3 20 C1-1,30CO- C1-13(CH2)4_ (S) -C4H3 Oa CH30CO- C1-13(CH24- (S) -C(CH3)202CO(CH2)2CH3 C21-150CO- C6H5CH2- (S) -CH2020CH3 CH30CO- C^(CHA2(S) _C4H502 b CH30CO- 2'C51-14N - (R( -CH502c 25 C21-150CO- X-C5H4N- (R) -CH(CH3)02CCH3 C21H150CO- X-C5H4N- (R) -CH(CH3)02COC2H5 C21-150CO4'_C51-i4N- (S) -CH202CC(CH3)3 CH30CO- 4'_C5H4N- (R) -CH(CH3)02CCH3 4-crotonolactonyl b-y-butyrolacton-4-yi '3-phthaildyl EXAMPLE 37

6P-Methylsu Ifonyloxymethyl pen icil la nic acid A. benzyl 6p-methylsulfonyloxymethylpenicillanate To a cooled (- 1 OOC.) solution of 800 mg. of benzy] 6p- hydroxymethylpenicillanate and 0.55 ml. of triethylamino in 25 mi. of methylene chloride was added 194 mg. of methylsulfonyl chloride. After one hour of stirring the reaction mixture was washed successively with water, water at pH 1.0, a saturated sodium 40 bicarbonate solution and a brine solution. The organic phase was dried over magnesium sulfate and evaporated to dryness to give 650 mg. of the desired product.

B. 6p-methylsulfonyloxymethylpenicillanic acid To a suspension of 300 mg. of 5% palladium-on-calcium carbonate, prereduced with hydrogen at47 psi for 45 min., in 20 m[. of methanol-water OA) was added 300 mg. of benzyi 6p- methyisuifonyloxymethyl penicillanate, and the reduction continued for 30 min at 47 psi. An additional 300 mg. of catalyst was added and the reduction continued for an additional 30 min. The spent catalyst was filtered and the methanol removed in vacuo from the filtrate. The aqueous residue was extracted with ethyl acetate and the pH of the aqueous adjusted to 2 with 6N hydrochloric acid. The acidified aqueous was extracted with fresh ethyl acetate and the organic layer separated and backwashed with a saturated brine solution. The organic layer was dried over magnesium sulfate and evaporated to give 269 mg. of the desired product as an oil.

The NMR spectrum (CIDC13) showed absorption at 1.56 (s, 3H), 1.68 (s, 3H), 3.06 (s, 3H), 4.1 (m, 1 H), 4.41 (s, 1 H), 4.52 (m, 2H), 5.47 (cl, 1 H, J=4Hz) and 8.3 (s, 1 H) ppm.

EXAMPLE 39

6p-[1 (S)Hydroxy-3-phenylpropyllpenicillanic acid To a suspension of 244 mg. of 5% palladium-on-calcium carbonate, prereduced with hydrogen at 50 psi for min., in 20 mi. of methanol-water (1: 1) was added 244 mg. of benzy] 6p- [1 (S(hydroxy)-3 phenylpropyllpenicillanate (Example 11 B), and the resulting mixture shaken in a hydrogen atmosphere at an initial pressure of 52 psi for one hour. At this time an-additional 244 mg. of catalyst was added and the hydrogenation continued for one hour. An addition 244 mg. of catalyst was again added and the reduction completed in one hour. The catalyst was filtered and' the methanol removed in vacuo. The aqueous residue was extracted with ethyl acetate and was then acidified to pH 1.8 with 6N hydrochloric acid. The acidified aqueous was extracted with fresh ethyl acetate, and the organic phase separated, backwashed with a brine solution and dried over magnesium sulfate. Removal of the solvent under reduced pressure gave 127 mg. of 35 the desired product, m.p. 135-137.51C.

The NMR spectrum (C13C13, DMSO-D6) showed absorption at 1.57 (s, 3H), 1. 59 (s, 3H), 1.8 (m, 2H), 2.8 (m, 2H), 3.6 (d, d, J=1 0, 4Hz), 4.0 (m, 1 H), 4.13 (s, 1 H), 5.4 (d, 1 H, J=41-1z) and 7.2 (s, 51-1) ppm.

1 EXAMPLE40 t 40 6p-[1 (R)Hydroxy-3-phenyl propyll pen icil la nic acid To a suspension of 369 mg. of 5% palladium-0n-calcium carbonate, prereduced with hydrogen at 52 psi for min, in 20 mi. of methanol-water (1: 1) was added 369 mg. of benzyl 6p-[1 (R)hydroxy-3 phenyl propyllpenicillanate (Example 11 B), and the resulting mixture shaken in a hydrogen atmosphere at 52 psi for one hour. Three 370 mg. portions of the catalyst were added every hour over a three hour period, followed by hydrogen at 52 psi. One hour after the last addition the catalystwas filtered and the methanol removed from the filtrate. The aqueous residue was extracted with ethyl acetate and then acidified to pH 1.5 with 6N hydrochloric acid. Fresh ethyl acetate was added to the acidified aqueous and the organic phase separated, washed with a brine solution and dried over magnesium sulfate. Removal of the solvent in vacuo gave 110 mg. of the desired product, m.p. 131-135'C.

The NMR spectrum (CIDC13, DiVISO-D6) showed absorption at 1.53 (s, 3H), 1. 65 (s, 3H), 1.8 (m, 2H), 2.8 (m, 2H), 3.5 (d, d, J =9,4Hz), 4.1 (m, 1 H), 4.3 (s, 1 H), 5.3 (d, 1 H, J-4Hz) and 7.3 (s, 51-1) ppm.

EXAMPLE 41 55 Pivaloyoxymethyl 6p-methylsulfonyloxymethylpenicillanate To a solution of 1.0 9. of 6p-methylsulfonyloxymethylpenicillanic acid sodium salt in 10 ml. of dimethylformamide cooled to 0-50C. is added 0.53 ml. of chloromethyl pivalate, and the resulting reaction mixture is allowed to stir at room temperature overnight. The mixture is poured into water-ethyl acetate, and the organic layer separated, backwashed with a brine solution and dried over magnesium sulfate. Removal of the solvent in vacuo gives the desired product.

GB 2 128 986 A EXAMPLE 38

Starting with the appropriate sulfonyl chloride and benzyl 6phydroxymethylpenicillanate and employing the procedure of Example 37, the following compounds are prepared:

H H 5 IPH3 S CH3 0 C02R 10 R (CH3)2CHSO2OCI-12 C1-13(C1-12)3SO2OCH2 C6H5SO2OCH2- 15 2-CH3C6H4SO2OCH2 4-CH30C61-14SO2OCH2 2-FC6H4S0p0CH2 2-CIC6H4SO2OCH2 2-BrC61-14SO2OCH2- 20 3-CF3C61-14SO2OCH2 4-CF3C6H4SO2OCI-12- t f 4 31 GB 2 128 986 A 31 EXAMPLE 42

Starting with the appropriate penicillanic acid and requisite halide, and employing the procedure of Example 41, the following compounds are prepared:

CH 3 S : CH 3 R N 0 11 CO 2 R 1 R R, 15 CH3SO2OCH2- -CH(CH3)02CCH3 CH3SO2OCH2- -CH202CC1-13 CH3S02OCH2- -CH202COCH(CH3)2 (C1-13)2CHS020CH2-CH202CC1-13 20 (CH3)2CHS020CH2- -CH202COCH3 (CH3)2CHCO2OCI-12- -CsH302a (C1-13)2CHS020CH2- -C(CH3)202CC(CH3)3 CH3WH2)3SO2OCI-12-CH(CH3)02CO(CH2)3CH3 -CH202CCH3 25 CH3WHA3SO2OCH2- b CH3(CH26SO2OCI-12-C41-1502 a C61-15C02OCH2- -C41-1302 C61-15SO2OCI-12- -CH(CH3)02COC2H5 C6H5SO2OCI-12- -CH202C(CH2)4CH3 2-CH3CrH4SO2OCH2- -CH202CC(CH3)3 30 2-CH3C6H4SO2OCI-12- -CH(CH3)02CC1-13 2-CH3C61-14SO3OCH2- -CH(CH3)02COC2H5 4-CH30C61-14SO2OCH2- -C81-1502c 4-CH30C6H4SO2OCI-12- -CH202CCH3 4-CH30C6H4SO2OCI-12- -CH202OCC(CH36 35 4-CH30C61-14SO2OCH2-CH(C1-13)02COC2H5 2-FC61-14SO2OCI-12- - C4H302 a 2-FC6H4S02.OCH2-CH202C(CHA4CH3 2-FC6H4SO2OCH2- -C(CH3)202(CHA2CH3 2-CIC61-14SO2OCH2-CH202COCH(CH3)2 40 2-C[C6H4SO2OCHP- -CH202CCH3 -CH(CH3)02COC2H5 2-CIC6H4SO2OCH2- a 2-CIC6H4SO2OCI-12- -C4H302 b 2-BrC61-14SO2OCH2- -C4H502 2-BrC6H4SO2OCH2- -CH202CCH3 45 2-BrC61-14SO2OCH2- -CH202COCH3 3-CF -CH202CC(CH3)3 3C6H4SO2OCH2- b 3-CF3Cr,H4SO2OCI-12- -C4H502 4-CF3C6H4SO2OCH2- -C(CH3)202CC(CH3)3 4-CF3C6H4SO2OCH2- -CH202CCH3 50 4-CF3C61-14SO2OCH2- -CH(CH3)02CO(CH2)3CH3 (S) C6H5(CH2)2CHOH- -CH202CC1-13 (S) C^(C1-12)2 CHOH- -CH(CH302COC2H5 (S) C61-15(CH2)2CHOH-CH202C(CH2)4CH3 (R) C6H5 (CH2)2CHOH- -C8H5 0 2 c 55 (R) C6H5 (CH2)2CH01-1- -C(CH3)202CO(CHA2CH3 (R) C^(CH2)2CHOH- -CH202COCH(C1-13)2 32 GB 2 128 986 A

Claims

1. A compound of the formula:

32 H H 5 - 1 CH 3 1 R S CH3 N 0 co 2 R 1 or a pharmaceutically acceptable base salt thereof, wherein R is 1- hydroxy-3-phenylpropyl and R, is benzyl, 15 hydrogen or an ester-forming residue readily hydrolyzable in vivo.

2. 6-p-(l-Hydroxy-3-phenylpropyl)pencillanic acid.

3. A pharamaceutical composition useful for treating bacterial infections in mammals, which comprises a pharmaceutically acceptable carrier, a Plactam antibiotic and a compound as claimed in claim 1 or claim 2 20 with the proviso that R, is not benzyl.

4. A pharmaceutical composition as claimed in claim 3, wherein R, is hydrogen, 1-(alkanoyloxy)ethyl having from four to seven carbon atoms, 1 methyl-l-(alkanoylloxy) ethyl having from five to eight carbon atoms, alkoxycarbonymethyl having from three to six carbon atoms, 1(alkoxycarbonyloxy)ethyl having from four to seven carbon atoms, 1-methyl1 -(alkoxycarbonyloxy)ethyl having from five to eight carbon 25 atoms, 3phtha I idyl, 4-croto nol actonyl or -y-butyrol acto n-4-yl.

Printed for Her Majesty's Stationery Office, by Croydon Printing Company Limited, Croydon. Surrey, 1984. Published by The Patent Office, 25 Southampton Buildings, London, WC2A lAY, from which copies may be obtained.

i r 40, c